In situ Studies of Crassulacean Acid Metobolism in Drymoglossum piloselloides, an Epiphytic Fern of the Humid Tropics

This paper reports autecological field-studies in Singaporeon Drymoglossum piloselloides (L.) Presl., an epiphytic fernof the humid tropics which is capable of performing Crassulaceanacid metabolism (CAM). As indicated by the gas exchange patternsand by the occurrence of a diurnal malic acid rhythm, the plantalso features CAM in situ at its natural sites. Both in well-wateredand in naturally droughted plants external CO₂ was taken upsolely during the night. Water stress decreased nocturnal CO₂uptake,but left the synthesis and storage of malic acid unaffected.This indicates that CO₂ recycling of respiratory CO₂ by CAMis ecophysiologically important at the high night temperaturestypical of the tropical habitats of the fern. The plants showeda diel fluctuation of cell-sap osmotic pressure which paralleledthat of malic acid, while the fluctuation of the xylem tensionfollowed the curve of transpiration more closely than it followedthat of the malic acid content. CAM in D. piloselloides wasclearly not limited by natural access to mineral ions and nitrogen.It is concluded that the ecophysiological advantage of CAM forD. piloselloides lies in a better water use efficiency as comparedwith C₃ ferns and in the salvaging of carbon by CO₂ recycling. Key words: CAM, epiphytic ferns, gas exchange, water relations     

Structural and metabolic properties of green tissue cultures from a CAM plant, Kalanchoë blossfeldiana hybr. Montezuma

Abstract Crassulacean acid metabolism (CAM) was studied in mixotrophic callus tissue cultures of Kalanchoë blossfeldiana hybr. Montezuma and compared with plants propagated from the calli. The ultrastructural properties of the green callus cells are similar to mesophyll cells of CAM plants except that occasionally abnormal mitochondria were observed. There was permanent net CO₂ output by the calli in light and darkness, which was lower in darkness than in light. The calli exhibited a diurnal rhythm of malic acid, with accumulation during the night and depletion during the day. ¹⁴C previously incorporated by dark CO₂ fixation into malate was transferred upon subsequent illumination into end products of photosynthesis. All these data indicate that CAM operates in the calli tissue. The results revealed that the capacity for CAM is obviously lower in the calli compared with plantlets developing from the calli, or with ‘adult’ plants. The data suggest also that CAM in the calli was not limited by the activities of CAM enzymes.     

Is Sedum acre L. a CAM plant?

Summary Sedum acre L. collected from its natural stands south of Darmstadt (Germany) showed ¹³C values typical for C₃ plants. This suggests that in situ at the natural stand CO₂ was fixed mainly via the C₃ mode of photosynthesis rather than via the CAM mode. However, experimental water stress shifts the CO₂ exchange pattern from the C₃ type to CAM type. Simultaneously, a diurnal rhythm of malic acid oscillation, typical for CAM, and increase of PEP-carboxylase and malic enzyme activities developed. Hence, Sedum acre is obviously to be classified as a facultative CAM plant. Because of the temperature characteristics of CO₂ exchange in Sedum acre, in situ CO₂ should be harvested from the atmosphere mainly during the seasons where water stress situations capable of inducing CAM are unlikely to occur.     

In situ studies on crassulacean acid metabolism in Sedum acre L. and Sedum mite Gil

Summary CO₂ exchange, the diurnal variations in the levels of malic, citric and isocitric acid, and the labelling pattern after ¹⁴CO₂ fixation were measured in Sedum acre and Sedum mite growing in situ. As predicted from laboratory experiments, drought changed the gas exchange pattern from a C₃ type to a crassulacean acid metabolism (CAM) type. This shift correlated with the development of a diurnal rhythm in the malic acid content. The results of ¹⁴CO₂ pulse-chase experiments suggest that in well-watered plants a CAM pattern of carbon flow already exists; hence water stress might enhance latent CAM rather than induce it. The in situ CAM performance by the Sedum species appeared to be highly susceptible to modulation by season and external factors, particularly light and temperature.CAM did not substantially contribute to total carbon gain in S. acre and S. mite. During most of their lifecycles the plants grow under conditions that favour CO₂ uptake by the C₃ pathway rather than by CAM. Hence, despite a capability to feature CAM, the ¹³C values found in S. acre and S. mite are those of C₃ plants.Abbreviations CAM Crassulacean Acid Metabolism - PEP-C Phosphoenolpyruvate-Carboxylase - DW Dry weight Dedicated to Prof. Dr. Dr. h.c. M. Evenari on the occasion of his 75th birthday and to Dr. K.F. Springer     

Studies on carbon flow in Crassulacean acid metabolism during the initial light period

In the Crassulacean acid metabolism (CAM) plants Kalanchoë tubiflora and Sedum morganianum a shift in the pathways occurs by which external CO₂ enters the metabolism during the initial light period (phase II of the diurnal CAM cycle). At the beginning of phase II, CO₂ is fixed mainly by the C₄ pathway; during late phase II, however, it is fixed mainly via the C₃ pathway. The C₃ pathway contributes to the phosphoenolpyruvate-carboxylase-mediated CO₂ fixation by the provision of three-carbon skeletons. Since the shift in the carbon-flow pathway is delayed after a CO₂-free night when malic-acid accumulation in the vacuoles is prevented, it is very likely that the amount of malic acid in the vacuole is integrated in the mechanism which controls CAM during the initial light period. A light-on signal at the beginning of phase II is not required to bring about the shifts in the carbon-flow pathways, as is shown by the reaction of plants to a prolonged dark period. A model of carbon flow during phase II is proposed.Abbreviations CAM Crassulacean acid metabolism - PEP-Case phosphoenolpyruvate carboxylase     

Effects of Various Levels of CO(2) on the Induction of Crassulacean Acid Metabolism in Portulacaria afra (L.) Jacq

In response to water stress, Portulacaria afra (L.) Jacq. (Portulacaceae) shifts its photosynthetic carbon metabolism from the Calvin-Benson cycle for CO₂ fixation (C₃) photosynthesis or Crassulacean acid metabolism (CAM)-cycling, during which organic acids fluctuate with a C₃-type of gas exchange, to CAM. During the CAM induction, various attributes of CAM appear, such as stomatal closure during the day, increase in diurnal fluctuation of organic acids, and an increase in phosphoenolpyruvate carboxylase activity. It was hypothesized that stomatal closure due to water stress may induce changes in internal CO₂ concentration and that these changes in CO₂ could be a factor in CAM induction. Experiments were conducted to test this hypothesis. Well-watered plants and plants from which water was withheld starting at the beginning of the experiment were subjected to low (40 ppm), normal (ca. 330 ppm), and high (950 ppm) CO₂ during the day with normal concentrations of CO₂ during the night for 16 days. In water-stressed and in well-watered plants, CAM induction as ascertained by fluctuation of total titratable acidity, fluctuation of malic acid, stomatal conductance, CO₂ uptake, and phosphoenolpyruvate carboxylase activity, remained unaffected by low, normal, or high CO₂ treatments. In well-watered plants, however, both low and high ambient concentrations of CO₂ tended to reduce organic acid concentrations, low concentrations of CO₂ reducing the organic acids more than high CO₂. It was concluded that exposing the plants to the CO₂ concentrations mentioned had no effect on inducing or reducing the induction of CAM and that the effect of water stress on CAM induction is probably mediated by its effects on biochemical components of leaf metabolism.     

Induction of a Crassulacean acid like metabolism in the C4 succulent plant,Portulaca oleracea L.: physiological and morphological changes are accompanied by specific modifications in phosphoenolpyruvate carboxylase

The induction of a Crassulacean acid like metabolism (CAM) was evidenced after 21–23 days of drought stress in the C₄ succulent plant Portulaca oleracea L. by changes in the CO₂ exchange pattern, in malic acid content and in titratable acidity during the day–night cycle. Light microscopy studies also revealed differences in the leaf structure after the drought treatment. Following the induction of the CAM-like metabolism, the regulatory properties of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), the enzyme responsible for the diurnal fixation of CO₂ in C₄ plants but nocturnal in CAM plants, were studied. The enzyme from stressed plants showed different kinetic properties with respect to controls, notably its lack of cooperativity, higher sensitivity to L-malate inhibition, higher PEP affinity and lower enzyme content on a protein basis. In both conditions, PEPC's subunit mass was 110 kDa, although changes in the isoelectric point and electrophoretic mobility of the native enzyme were observed. In vivo phosphorylation and native isoelectrofocusing studies indicated variations in the phosphorylation status of the enzyme of samples collected during the night and day, which was clearly different for the control and stressed groups of plants. The results presented suggest that PEPC activity and regulation are modified upon drought stress treatment in a way that allows P. oleracea to perform a CAM-like metabolism. This revised version was published online in August 2006 with corrections to the Cover Date.     

Seasonal diurnal acid rhythms in two aquatic crassulacean acid metabolism plants

Summary The diurnal change in titrable acidity in two aquatic CAM plants, Littorella uniflora var. americana (Fern.) Gl. and Isoetes macrospora Duriev., growing at two sites, was monitored at biweekly intervals for two years during the ice-free period. Both plants exhibited the classic pattern of CAM acitivity, with deacidification 60 to 90% complete by noon. The maximum diurnal acid rhythms observed were 169±7, and 154±20 eq·g^-1 fresh weight for the two Littorella populations, and 182±9 and 133±16 eq·g^-1 fresh weight for the two Isoetes populations. The seasonal pattern of the diurnal acid rhythm was correlated with temperature and light. The maximum activity occurred in midsummer, with negligible activity under ice cover. This pattern was similar to that of terrestrial CAM plants from non-arid environments. Comparison of CAM activity for populations of the same species indicates that the magnitude of CAM activity is closely related to plant productivity, and appears to be related to light and perhaps CO₂ availability. In these plants, CAM extends the diel period of carbon accumulation and contributes 40 to 50% of the annual carbon gain. The prolonged period of carbon acquisition and effective conservation of respired CO₂ via CAM is of paramount importance in the growth and productivity of these plants in oligotrophic environments.     

Differential effects of drought and light levels on accumulation of citric and malic acids during CAM in Clusia

Night-time citrate accumulation has been proposed as a response to stress in CAM plants. To address this hypothesis, gas exchange patterns and nocturnal acid accumulation in three species of Clusia were investigated under controlled conditions with regard to water stress and responses to low and high photosynthetic photon flux density (PPFD). Under high PPFD, leaves of Clusia nocturnally accumulated large amounts of both malic and citric acids. Under low PPFD and well-watered conditions, substantial night-time citrate accumulation persisted, whereas malate accumulation was close to zero. Malate accumulation and night-time CO₂ uptake from the atmosphere declined in all three species during prolonged drought periods, whereas citrate accumulation remained similar or increased. Recycling of respiratory CO₂ was substantial for both well-watered and water-stressed plants. The suggestion that citrate accumulation is energetically more favourable than malate accumulation is not supported if the source of CO₂ for the formation of malate is respiratory CO₂. However, the breakdown of citric acid to pyruvate in the light period releases three molecules of CO₂, while the breakdown of malic acid releases only one CO₂ per pyruvate formed. Thus, citric acid should be more effective than malic acid as a mechanism to increase CO₂ concentration in the mesophyll and may help to prevent photoinhibition. Organic acid accumulation also affected the vacuolar pH, which reached values of 2·6–3·0 at dawn. At these pH values, the transport of 2H⁺/ATP is still feasible, suggesting that it is the divalent form of citrate which is being transported in the vacuoles. Since citrate is a well-known buffer, and Clusia spp. show the largest day-night changes in organic acid levels measured in any CAM plant, it is possible that citrate increases the buffer capacity of the vacuoles. Indeed, malate and titratable acidity levels are positively related to citrate levels. Moreover, Clusia species that show the highest nocturnal accumulation of organic acids are also the ones that show the greatest changes in citric acid levels.     

Fluorescence Quenching in the Varied Photosynthetic Modes of Portulacaria afra (L.) Jacq

The kinetics of chlorophyll fluorescence were measured in Portulacaria afra (L.) Jacq. when the plants were functioning in either Crassulacean acid metabolism (CAM) or C₃/CAM cycling (called cycling) modes, as determined by fluctuation in titratable acidity and gas exchange properties. Cycling plants showed primarily daytime CO₂ uptake typical of C₃ plants, but with a slight diurnal acid fluctuation, whereas CAM plants showed nocturnal CO₂ uptake, daytime stomatal closure, and a large diurnal acid fluctuation. Results from fluorescence measurements indicated no significant differences in photochemical quenching between cycling and CAM plants; however, sizable differences were detected in nonphoto-chemical quenching (q_n), with the largest differences being observed during the middle of the day. Cycling plants had lower q_n than CAM plants, indicating altered photosynthetic regulation processes. This q_n difference was believed to be related to reduced internal CO₂ concentration in the CAM plants because of daytime stomatal closure and reduced deacidification rates in the late afternoon when most of the malic acid has been utilized. Experimentally, higher external CO₂ given to plants in the CAM mode resulted in a decline in q_n in comparison to that measured in plants in the cycling mode. No changes were observed in photochemical quenching when CO₂ was added.     

Correlation between CAM-Cycling and Photosynthetic Gas Exchange in Five Species of Talinum (Portulacaceae)

Photosynthetic gas exchange and malic acid fluctuations were monitored in 69 well-watered plants from five morphologically similar species of Talinum in an investigation of the ecophysiological significance of the Crassulacean acid metabolism (CAM)-cycling mode of photosynthesis. Unlike CAM, atmospheric CO₂ uptake in CAM-cycling occurs exclusively during the day; at night, the stomata are closed and respiratory CO₂ is recaptured to form malic acid. All species showed similar patterns of day-night gas exchange and overnight malic acid accumulation, confirming the presence of CAM-cycling. Species averages for gas exchange parameters and malic acid fluctuation were significantly different such that the species with the highest daytime gas exchange had the lowest malic acid accumulation and vice versa. Also, daytime CO₂ exchange and transpiration were negatively correlated with overnight malic acid fluctuation for all individuals examined together, as well as within one species. This suggests that malic acid may effect reductions in both atmospheric CO₂ uptake and transpiration during the day. No significant correlation between malic acid fluctuation and water-use efficiency was found, although a nonsignificant trend of increasing water-use efficiency with increasing malic acid fluctuation was observed among species averages. This study provides evidence that CO₂ recycling via malic acid is negatively correlated with daytime transpirational water losses in well-watered plants. Thus, CAM-cycling could be important for survival in the thin, frequently desiccated soils of rock outcrops on which these plants occur.     

Stomatal responses to changes in air humidity are not necessarily linked to nocturnal CO2 uptake in the CAM plant Plectranthus marrubioides Benth. (Lamiaceae)

Plants of the crassulacean acid metabolism (CAM) species Plectranthus marrubioides (Lamiaceae) were subjected to short- and long-term changes in air humidity in controlled-environment experiments. Stomata of well-watered individuals of this all-cell leaf-succulent taxon responded directly, quickly and reversibly to variations of the water vapour gradient between leaf and air (Δw). Mean night-time leaf conductance to water vapour decreased curvilinearly with increasing Δw but linearly with lowered relative air humidity. Stomatal response was generally independent of the prevailing temperature and was not linked to CO₂ uptake rates. Therefore, net night-time carbon gain, nocturnal malic acid accumulation and, thus, relative carbon recycling were not influenced by changes in air humidity in the temperature range tested. Mean nocturnal molar water use efficiency, however, decreased with decreasing air humidity because of the increased transpirational water loss. If watering was repeatedly withheld for several days during the experiments, employing a temperature regime of 35/30°C day and night, stomatal conductance became low enough to inhibit CO₂ uptake, but only at the highest Δw. The results suggest that drought stress was necessary to increase responsiveness of plants to the point where CAM was also inhibited by decreases in air humidity.     

Comparative ecophysiology of CAM and C3 bromeliads. III. Environmental influences on CO2 assimilation and transpiration

Abstract Field measurements of the gas exchange of epiphytic bromeliads were made during the dry season in Trinidad in order to compare carbon assimilation with water use in CAM and C₃ photosynthesis. The expression of CAM was found to be directly influenced by habitat and microclimate. The timing of nocturnal CO₂ uptake was restricted to the end of the dark period in plants found at drier habitats, and stomatal conductance in two CAM species was found to respond directly to humidity or temperature. Total night-time CO₂ uptake, when compared with malic-acid formation (measured as the dawn-dusk difference in acidity, ΔH⁺), could only account for 10–40% of the total ΔH⁺ accumulated. The remaining malic acid must have been derived from the refixation of respired CO₂ (recycling). Within the genus Aechmea (12 samples from four species), recycling was significantly correlated with night temperature at the six sample sites. Recycling was lowest in A. fendleri (54% of ΔH⁺ derived from respired CO₂), a CAM bromeliad with little water-storage parenchyma that is restricted to wetter, cooler regions of Trinidad. Gas-exchange rates of C₃ bromeliads were found to be similar to those of the CAM bromeliads, with CO₂ uptake from 1 to 3 μmol m^?2 s^?1 and stomatal conductances generally up to 100 mmol m^?2 s^?1. The midday depression of photosynthesis occurred in exposed habitats, although photosynthetically active radiation (PAR) limited photosynthesis in shaded habitats. CO₂ uptake of the C₃ bromeliad Guzmania lingulata was saturated at around 500 μmol m^?2 s^?1 PAR, suggesting that epiphytic plants found in the shaded forest understorey are shade-tolerant rather than shade-demanding. Transpiration ratios (TR) during CO₂ fixation in CAM (Phase I and IV) and C₃ bromeliads were compared at different sites in order to assess the efficiency of water utilization. For the epiphytes displaying marked uptake of CO₂, TR were found to be lower than many previously published values. In addition, the average TR values were very similar for dark CO₂ uptake in CAM (42 ± 41, n= 12), Phase IV of CAM (69 ± 36, n= 3) and for C₃ photosynthesis (99 ± 73, n= 4) in these plants. It appears that recycling of respired CO₂ by CAM bromeliads and efficient use of water in all phases of CO₂ uptake are physiological adaptations of bromeliads to arid microclimates in the humid tropics.     

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.     

Diurnal changes in volume and specific tissue weight of crassulacean Acid metabolism plants

The diurnal variations in volume and in specific weight were determined for green stems and leaves of Crassulacen acid metabolism (CAM) plants. Volume changes were measured by a water displacement method. Diurnal variations occurred in the volume of green CAM tissues. Their volume increased early in the light period reaching a maximum about mid-day, then the volume decreased to a minimum near midnight. The maximum volume increase each day was about 2.7% of the total volume. Control leaves of C₃ and C₄ plants exhibited reverse diurnal volume changes of 0.2 to 0.4%. The hypothesis is presented and supported that green CAM tissues should exhibit a diurnal increase in volume due to the increase of internal gas pressure from CO₂ and O₂ when their stomata are closed. Conversely, the volume should decrease when the gas pressure is decreased.

The second hypothesis presented and supported was that the specific weight (milligrams of dry weight per square centimeter of green surface area) of green CAM tissues should increase at night due to the net fixation of CO₂. Green CAM tissues increased their specific weight at night in contrast to control C₃ and C₄ leaves which decreased their specific weight at night. With Kalanchoë daigremontiana leaves, the calculated increase in specific leaf weight at night based on estimates of carbohydrate available for net CO₂ fixation was near 6% and the measured increase in specific leaf weight was 6%.

Diurnal measurements of CAM tissue water content were neither coincident nor reciprocal with their diurnal patterns of either volume or specific weight changes.

     

Le vie fotosintetiche C3, C4 e CAM nel contesto ecologico

Abstract

Ecological aspects of C₃, C₄ and CAM photosynthetic pathways. - Three different photosynthetic CO₂ fixation pathways are known to occur in higher plants. However all three pathways ultimately depend on the Calvin-Benson cycle for carbon reduction. The oxygenase activity of RuBP carboxilase is responsible for photorespiratory CO₂ release. Both C₄ and CAM pathways behave as a CO₂ concentrating mechanism which prevent photorespiration. The CO₂-concentrating mechanism in C₄ plants is based on intracellular symplastic transport of C₄ dicarboxylic acids from mesophyll-cells to the adjacent bundle-sheath cells. On the contrary in CAM plants the CO₂-concentrating mechanism is based on the intracellular transport of malic acid into and out of the vacuole.

The C₄ photosynthetic pathway as compared to the C₃ pathway permits higher rates of CO₂ fixation in high light and high temperature environments at low costs in terms of water loss, given the stability of the photosynthetic apparatus under such conditions.

CAM is interpreted as an adaptation to arid environments because it enables carbon assimilation to take place at very low water costs during the night when the evaporative demand is low. Nevertheless many aquatic species of Isoetes and some relatives are CAM, suggesting the adaptive role of CAM to environments which become depleted in CO₂.

The photosynthetic carbon fixation pathway certainly contributes to the ecological success of plants in different environments. However the distribution of plants may also reflect their biological history. On the other hand plants with different photosynthetic pathways coexist in many communities and tend to share resources in time. In any case some generalizations are possible: C₄ plants enjoy an ecological advantage in hot, moist, high light regions while the majority of species in desert environments are C₃; CAM plants are more frequent in semiarid regions with seasonal rainfall, coastal fog deserts, and in epiphytic habitats in tropical rain forests.     

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.

     

Crassulacean acid metabolism photosynthesis: `working the night shift'

Crassulacean acid metabolism (CAM) can be traced from Roman times through persons who noted a morning acid taste of some common house plants. From India in 1815, Benjamin-Heyne described a `daily acid taste cycle' with some succulent garden plants. Recent work has shown that the nocturnally formed acid is decarboxylated during the day to become the CO₂ for photosynthesis. Thus, CAM photosynthesis extends over a 24-hour day using several daily interlocking cycles. To understand CAM photosynthesis, several landmark discoveries were made at the following times: daily reciprocal acid and carbohydrate cycles were found during 1870 to 1887; their precise identification, as malic acid and starch, and accurate quantification occurred from 1940 to 1954; diffusive gas resistance methods were introduced in the early 1960s that led to understanding the powerful stomatal control of daily gas exchanges; C₄ photosynthesis in two different types of cells was discovered from 1965 to ∼1974 and the resultant information was used to elucidate the day and night portions of CAM photosynthesis in one cell; and exceptionally high internal green tissue CO₂ levels, 0.2 to 2.5%, upon the daytime decarboxylation of malic acid, were discovered in 1979. These discoveries then were combined with related information from C₃ and C₄ photosynthesis, carbon biochemistry, cellular anatomy, and ecological physiology. Therefore by ∼1980, CAM photosynthesis finally was rigorously outlined. In a nutshell, 24-hour CAM occurs by phosphoenol pyruvate (PEP) carboxylase fixing CO₂(HCO₃ ⁻) over the night to form malic acid that is stored in plant cell vacuoles. While stomata are tightly closed the following day, malic acid is decarboxylated releasing CO₂ for C₃ photosynthesis via ribulose bisphosphate carboxylase oxygenase (Rubisco). The CO₂ acceptor, PEP, is formed via glycolysis at night from starch or other stored carbohydrates and after decarboxylation the three carbons are restored each day. In mid to late afternoon the stomata can open and mostly C₃ photosynthesis occurs until darkness. CAM photo-synthesis can be both inducible and constitutive and is known in 33 families with an estimated 15 to 20 000 species. CAM plants express the most plastic and tenacious photosynthesis known in that they can switch photosynthesis pathways and they can live and conduct photosynthesis for years even in the virtual absence of external H₂O and CO₂, i.e., CAM tenaciously protects its photosynthesis from both H₂O and CO₂ stresses. This revised version was published online in August 2006 with corrections to the Cover Date.     

ISOETES HOWELLII: A SUBMERGED AQUATIC CAM PLANT?

In the leaves (but not corms) of the submerged aquatic plant Isoetes howellii, malic acid concentration fluctuates from 1–3 mg g^–1 FW in the evening to 7–13 mg g^–1 FW in the morning. Associated with this is a change in pH (a.m. pH 3–4 vs. p.m. pH 5–6) and titratable acidity (75–200 μ eq g^–1 FW change in acidity between morning and evening) of the plant extract. ¹⁴CO₂-fixation experiments indicate that carbon is fixed in both the light and the dark, though the amount of carbon fixed in the light is more than double that fixed in the dark. Autoradiographs show 89% of dark-fixed CO₂ ends up in malic acid and the remainder in citric acid, whereas these two acids constitute less than 5% of the light-fixation products. It is suggested that CAM metabolism in this aquatic species may be related to the lower availability of CO₂ for photosynthesis during the day than during the night in its aquatic environment.     

Evidence that the induction of crassulacean acid metabolism by water stress in Mesembryanthemum crystallinum (L.) involves root signalling

The aim of this study was to investigate whether the root system of Mesembryanthemum crystallinum (L.) plays a role in triggering the induction of crassulacean acid metabolism (CAM) during water stress. Depriving well-irrigated plants of water, by allowing the soil surrounding the roots to dry, caused increased daily losses in leaf relative water content (RVVC) and mesophyll cell turgor pressure. The RWC of the roots also declined. Subsequently plants exhibited physiological characteristics of CAM photosynthesis (i.e. diurnal fluctuations in leaf titratable acidity and nocturnal net CO₂ fixation). When the root system of plants was divided equally between two soil compartments and one half deprived of water, plants exhibited physiological characteristics of CAM without prior changes in leaf RWC content or mesophyll cell turgor pressure. Only the RWC of the water-stressed portion of the roots was reduced. These data suggest that in water-stressed plants daily changes in leaf water relations greater than those observed in well-irrigated plants, are not essential to trigger CAM expression. It is probable that a reduction in soil water availability can be perceived by the roots of M. crystallinum and that this information is conveyed to the leaves triggering the transition from C₃ to CAM photosynthesis.