Crassulacean Acid Metabolism Activity Research Articles

Epiphytic CAM bromeliads indicate vulnerability of tropical forest communities to climate change.

Vascular epiphytes have a variety of mechanisms to trap and retain water, including crassulacean acid metabolism (CAM). Niche segregation was investigated for epiphytic bromeliads on the tropical Caribbean island of Trinidad, where habitats range from lowland deciduous forests to high-rainfall montane tropical forests, ~1000 m in elevation. Four tank-impounding bromeliad epiphytes in the genus Aechmea (Ae. aquilega, Ae. fendleri, Ae. nudicaulis and Ae. dichlamydea) with CAM were mapped across their distinct geographical and elevational zonations in northern Trinidad and Tobago. Species distribution modelling was used to determine environmental limitations for each species. Anatomical and physiological measurements included leaf succulence traits, gas exchange and CAM activity; hydraulic conductance and vulnerability; stomatal sensitivity and quantum yield responses to nocturnal temperature and long-term water deficits. A total of 2876 field observations identified the transitions between the lowland Ae. aquilega and montane Ae. fendleri, occurring >500 m a.s.l. at the drier western end of the Northern Mountain Range and at progressively lower elevations towards the wetter, eastern region. Anatomical and physiological sensitivities of gas exchange, CAM activity and water use, and responses to elevated nocturnal temperatures and drought, were markedly different for Ae. fendleri compared with Ae. aquilega or the ubiquitous Ae. nudicaulis. The species distribution model highlighted the susceptibility of Ae. fendleri to a changing climate. For each species, physiological and anatomical traits were tailored to environmental tolerances, consistent with specialist or generalist niche preferences. Using Intergovernmental Panel on Climate Change scenarios, we predict that rapid rainfall and temperature changes will lead to the loss of Ae. fendleri and associated lower (and upper) montane forest communities from Trinidad, seriously impacting both biodiversity and critical ecosystem functions here and in other tropical island habitats. Epiphytic bromeliads act as markers for threatened communities, and their physiological tolerances represent key indicators of climate change impacts.

The CAM lineages of planet Earth.

The growth of experimental studies of crassulacean acid metabolism (CAM) in diverse plant clades, coupled with recent advances in molecular systematics, presents an opportunity to re-assess the phylogenetic distribution and diversity of species capable of CAM. It has been more than two decades since the last comprehensive lists of CAM taxa were published, and an updated survey of the occurrence and distribution of CAM taxa is needed to facilitate and guide future CAM research. We aimed to survey the phylogenetic distribution of these taxa, their diverse morphology, physiology and ecology, and the likely number of evolutionary origins of CAM based on currently known lineages. We found direct evidence (in the form of experimental or field observations of gas exchange, day-night fluctuations in organic acids, carbon isotope ratios and enzymatic activity) for CAM in 370 genera of vascular plants, representing 38 families. Further assumptions about the frequency of CAM species in CAM clades and the distribution of CAM in the Cactaceae and Crassulaceae bring the currently estimated number of CAM-capable species to nearly 7 % of all vascular plants. The phylogenetic distribution of these taxa suggests a minimum of 66 independent origins of CAM in vascular plants, possibly with dozens more. To achieve further insight into CAM origins, there is a need for more extensive and systematic surveys of previously unstudied lineages, particularly in living material to identify low-level CAM activity, and for denser sampling to increase phylogenetic resolution in CAM-evolving clades. This should allow further progress in understanding the functional significance of this pathway by integration with studies on the evolution and genomics of CAM in its many forms.

Morphological, photosynthetic, and CAM physiological responses of the submerged macrophyte Ottelia alismoides to light quality

Light quality strongly affect a great many physiological processes in terrestrial plants. Whatever, few studies have investigated how the submerged macrophytes response to different light quality. The present research aimed to understand how light quality affect morphogenesis, photosynthesis, as well as crassulacean acid metabolism (CAM) operation during the growth of Ottelia alismoides , a freshwater facultative CAM macrophyte. The plants were grown under three light qualities of white (W), red (R) and blue (B) lights with same light intensity of 100 μmol m −2 s −1 . Leaf development, leaf anatomic characteristics, photosynthetic traits, as well as CAM metabolism were determined on the newly produced leaves during light treatment. Monochromatic R stimulated leaf blade and petiole elongation, leaf area and dry weight, but reduced leaf thickness, negatively influenced chlorophyll content and Fv/Fm. On the contrary, B- grown plants were dwarf and with compact size as well as lower leaf dry weight. The content of photosynthetic product starch under B was significantly lower as compared with R, moreover, combined with the analysis of ultrastructural characteristics of epidermal and mesophyll chloroplasts, it showed that B inhibited starch to be translocated out of epidermal chloroplasts. Furthermore, both R- and B- grown plants exhibited obvious diurnal acidity fluctuation comparable to W- grown plants, and had high activity of enzymes including ribulose-bisphosphate carboxylase/oxygenase (Rubisco), phosphoenolpyruvate carboxylase (PEPC) and pyruvate phosphate dikinase (PPDK) that required for CAM cycle, indicating that CAM was present in both R- and B- grown plants and monochromatic R and B did not affect CAM activity. In conclusion, monochromatic red and blue light differentially played their special roles in regulating leaf growth and morphogenesis, as well as photosynthetic metabolism in O. alismoides . • O. alismoides displayed strong morphological plasticity to light quality. • Leaf elongation was promoted by red light but inhibited by blue light. • Red light favoured O. alismoides leaf development and photosynthesis. • Blue light inhibited translocation of starch out of epidermal chloroplasts. • O. alismoides exhibited comparable CAM activity under white, red and blue light.

Evolutionary history of CAM photosynthesis in NeotropicalClusia: insights from genomics, anatomy, physiology and climate

AbstractClusia is a remarkable genus of Neotropical woody plants as its members engage in either C3 photosynthesis or employ, to varying degrees, crassulacean acid metabolism (CAM) photosynthesis. Information about the evolutionary history of CAM in Clusia is scarce. Restriction site-associated sequencing of 64 species (20% of the genus) provided strong support for most of the previously recognized nine lineages. Ancestral reconstruction using maximum parsimony or maximum likelihood under a one-rate model suggested that CAM evolved at least four times independently from a most recent common ancestor (MRCA) with C3, whereas a maximum likelihood two-rate model suggested that CAM was already present in the MRCA followed by reversions to C3 in several lineages. Phylogenetic generalized least square analysis assessed variation in seven leaf anatomical traits and CAM activity measured as δ 13C. Results indicate that CAM is highly correlated with palisade mesophyll layer thickness and cell size. In addition, correlation between 19 bioclimatic variables and δ 13C was evaluated. It was found that CAM is positively correlated with habitats with a more severe dry season and greater precipitation seasonality. Since CAM is weakly and/or only periodically expressed in many Clusia spp., and thus not readily reflected in δ 13C, future analysis of phylogenetically-informed CAM expression in Clusia must include physiological measurements such as CO2 exchange and/or diel changes in leaf acidity for each species under investigation.

Seasonal changes in photosynthesis for the epiphytic bromeliad Tillandsia brachycaulos in a tropical dry deciduous forest

Background: Sunlight stress and drought affect plants by inducing various biochemical and physiological responses, which reduce growth. Seasonal changes in light and water availability that occur in forest canopies, where epiphytes occur, are extreme.
 Questions: What are the seasonal changes in photosynthesis for an abundant epiphytic bromeliad in contrasting microenvironments? Is Crassulacean acid metabolism (CAM) an important feature of photoprotection for this epiphyte?
 Studied species: Tillandsia brachycaulos Schltdl. (Bromeliaceae)
 Study site and dates: Canopy of the tropical dry deciduous forest of Dzibilchaltún National Park, Yucatan, Mexico during the rainy season 2008 and dry season 2009.
 Methods: Diurnal measurements of photosystem II efficiency, titratable acidity, leaf water potential, and photosynthetic pigment concentration were measured during the dry and rainy seasons in adult plants of T. brachycaulos in shaded and exposed microenvironments. The prevailing environmental conditions (photon flux density, precipitation, air temperature and relative humidity) were also seasonally characterized.
 Results: The highest irradiance occurred during the dry season caused photo-inactivation, a decrease of the quantum efficiency of photosystem II and a reduction in CAM activity of about 40 % in leaves of exposed plants of T. brachycaulos. During the rainy season, the leaf water potential of exposed and shaded plants of T. brachycaulos was lower at midday than at predawn, indicating water loss during the day.
 Conclusions: Individuals of T. brachycaulos reduced CAM activity during the dry season; and, during the rainy season, increased carbon gain by stomata opening during phase II and IV of CAM.

The Development of Crassulacean Acid Metabolism (CAM) Photosynthesis in Cotyledons of the C4 Species, Portulaca grandiflora (Portulacaceae).

Portulaca grandiflora simultaneously utilizes both the C4 and Crassulacean acid metabolism (CAM) photosynthetic pathways. Our goal was to determine whether CAM developed and was functional simultaneously with the C4 pathway in cotyledons of P. grandiflora. We studied during development whether CAM would be induced with water stress by monitoring the enzyme activity, leaf structure, JO2 (rate of O2 evolution calculated by fluorescence analysis), and the changes in titratable acidity of 10 and 25 days old cotyledons. In the 10 days old cotyledons, C4 and CAM anatomy were evident within the leaf tissue. The cotyledons showed high titratable acid levels but a small CAM induction. In the 25 days old cotyledons, there was a significant acid fluctuation under 7 days of water stress. The overall enzyme activity was reduced in the 10 days old plants, while in the 25 days old plants CAM activity increased under water-stressed conditions. In addition to CAM, the research showed the presence of glycine decarboxylase in the CAM tissue. Thus, it appears both pathways develop simultaneously in the cotyledons but the CAM pathway, due to anatomical constraints, may be slower to develop than the C4 pathway. Cotyledons showed the ancestral Atriplicoid leaf anatomy, which leads to the question: Could a CAM cell be the precursor to the C4 pathway? Further study of this may lead to understanding into the evolution of C4 photosynthesis in the Portulaca.

What does the RuBisCO activity tell us about a C3-CAM plant?

Plants that perform the Crassulacean acid metabolism (CAM), which obtain CO2 overnight and convert it mainly in malic acid, successfully grow in environments with water and nutrient shortages, that is partly associated with their higher water- and nitrogen-use efficiencies. Water and nutrient limitations can impair photosynthesis through the reduction of RuBisCO and increment of photorespiration, disturbing the plant carbon balance. In this context, we conducted a controlled experiment with the epiphytic C3-CAM bromeliad Guzmania monostachia to investigate how the combined water and nutritional deficits affect the activity of RuBisCO and its activation state (RAS), and to evaluate the efficiency of photosynthesis during the transition from C3 to CAM. Apart from an increase in CAM activity, bromeliads submitted to both water and nutritional deficits showed higher RAS values and unaltered RuBisCO activity compared to C3 bromeliads and, surprisingly, the maximum quantum efficiency of photosynthesis increased. Glucose, fructose and starch levels were maintained, while sucrose concentrations increased over time. These results, combined with the high RAS values, suggest an increased efficiency of RuBisCO functioning. Our results reinforce the ability of epiphytic bromeliads to deal with stressful habitats by a higher efficiency of RuBisCO during the transition to CAM, another feature that may allow their evolution in the epiphytic environment.

The Analysis of Leaf Traits of Eight Ottelia Populations and Their Potential Ecosystem Functions in Karst Freshwaters in China.

Submerged macrophytes play a structuring role in the shallow freshwater ecosystem by increasing the heterogeneous state in freshwaters. The macrophytes in genus Ottelia were featured for their broad leaves, which might consequently produce specialized functions that differed from other submerged species. To explore the potential ecological role of Ottelia, a field investigation was conducted on leaf traits in eight populations of Ottelia ranging from the southwestern Yunnan–Guizhou plateau to the southern Hainan island in China covering a distance of >1,700 km. The eight populations included all the extant Ottelia species and varieties in China except the well-documented O. alismoides. Carbon-related traits [bicarbonate usage, photosynthetic characteristics, capability of Crassulacean acid metabolism (CAM)], pigment content and parameters of chlorophyll fluorescence, morphology and mass of the leaves were determined. The different populations showed distinct functional traits of mature leaves; O. acuminata var. songmingensis had the thickest and longest leaf with CaCO3 precipitation on the both sides of the leaf, and O. cordata showed putative CAM activity with the highest diel acidity changes 12.5 μequiv g-1 FW. Our results indicated an important role of Ottelia populations in carbon cycling as the dominant species in karst freshwaters in China.

Different CO2 acclimation strategies in juvenile and mature leaves of Ottelia alismoides.

The freshwater macrophyte, Ottelia alismoides, is a bicarbonate user performing C4 photosynthesis in the light, and crassulacean acid metabolism (CAM) when acclimated to low CO2. The regulation of the three mechanisms by CO2 concentration was studied in juvenile and mature leaves. For mature leaves, the ratios of phosphoenolpyruvate carboxylase (PEPC) to ribulose-bisphosphate carboxylase/oxygenase (Rubisco) are in the range of that of C4 plants regardless of CO2 concentration (1.5-2.5 at low CO2, 1.8-3.4 at high CO2). In contrast, results for juvenile leaves suggest that C4 is facultative and only present under low CO2. pH-drift experiments showed that both juvenile and mature leaves can use bicarbonate irrespective of CO2 concentration, but mature leaves have a significantly greater carbon-extracting ability than juvenile leaves at low CO2. At high CO2, neither juvenile nor mature leaves perform CAM as indicated by lack of diurnal acid fluctuation. However, CAM was present at low CO2, though the fluctuation of titratable acidity in juvenile leaves (15-17µequiv g-1 FW) was slightly but significantly lower than in mature leaves (19-25µequivg-1 FW), implying that the capacity to perform CAM increases as leaves mature. The increased CAM activity is associated with elevated PEPC activity and large diel changes in starch content. These results show that in O. alismoides, carbon-dioxide concentrating mechanismsare more effective in mature compared to juvenile leaves, and C4 is facultative in juvenile leaves but constitutive in mature leaves.

Ammonium intensifies CAM photosynthesis and counteracts drought effects by increasing malate transport and antioxidant capacity in Guzmania monostachia.

Guzmania monostachia (Bromeliaceae) is a tropical epiphyte capable of up-regulating crassulacean acid metabolism (CAM) in its photosynthetic tissues in response to changing nutrient and water availability. Previous studies have shown that under drought there is a gradient of increasing CAM expression from the basal (youngest) to the apical (oldest) portion of the leaves, and additionally that nitrogen deficiency can further increase CAM intensity in the leaf apex of this bromeliad. The present study investigated the inter-relationships between nitrogen source (nitrate and/or ammonium) and water deficit in regulating CAM expression in G. monostachia leaves. The highest CAM activity was observed under ammonium nutrition in combination with water deficit. This was associated with enhanced activity of the key enzyme phosphoenolpyruvate carboxylase, elevated rates of ATP- and PPi-dependent proton transport at the vacuolar membrane in the presence of malate, and increased transcript levels of the vacuolar malate channel-encoding gene, ALMT. Water deficit was consistently associated with higher levels of total soluble sugars, which were maximal under ammonium nutrition, as were the activities of several antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase). Thus, ammonium nutrition, whilst associated with the highest degree of CAM induction in G. monostachia, also mitigates the effects of water deficit by osmotic adjustment and can limit oxidative damage in the leaves of this bromeliad under conditions that may be typical of its epiphytic habitat.

Antioxidant capacity along the leaf blade of the C3-CAM facultative bromeliad Guzmania monostachia under water deficit conditions.

Guzmania monostachia (L.) Rusby ex Mez is an epiphytic, rosette-shaped bromeliad that displays variable degrees of crassulacean acid metabolism (CAM) along the leaf under water deficit. The aim of our study was to evaluate whether the production-scavenging system of reactive oxygen species (ROS) along the leaf length of G. monostachia plants is related to the foliar gradient of CAM when irrigation is withheld. Among the leaf portions, the apex was exposed to the highest photosynthetic flux density and presented the highest relative water content, CAM activity, hydrogen peroxide and lipid peroxidation after treatment. Hence, the most intense CAM at the leaf apex may not have prevented higher oxidative burden in that region during water deficiency. However, the photosynthetic efficiency in the apex seemed unaffected by irrigation withholding or light intensity. The leaf apex also had the highest carotenoid content and increased superoxide dismutase and ascorbate peroxidase activities under treatment. Hence, G. monostachia was able to maintain ROS under tolerable levels by increasing antioxidant capacity. Our results suggest the metabolic differences within the same leaf under water deficit may derive from the gradient of light incidence, which emphasises the physiological plasticity this bromeliad applies to adapt to the adverse conditions of the canopy.

Adjustments in CAM and enzymatic scavenging of H2O2 in juvenile plants of the epiphytic bromeliad Guzmania monostachia as affected by drought and rewatering

Juvenile plants of epiphytes such as bromeliads are highly prone to dehydration under drought conditions. It is likely that young epiphytes evolved mostly metabolic strategies to resist drought, which may include the plastic modulation of the enzymatic antioxidant system and crassulacean acid metabolism (CAM). Few studies have investigated such strategies in juvenile epiphytes, although such research is important to understand how these plants might face drought intensification derived from potential climatic alterations. The epiphytic CAM bromeliad Guzmania monostachia (L.) Rusby ex Mez var. monostachia is known to have plastic responses to drought, but no reports have focused on the metabolism of juvenile plants to drought and recovery. Hence, we aimed to verify how juvenile G. monostachia plants adjust malate (indicative of CAM), H2O2 content and enzymatic scavenging in response to drought (eight days without irrigation) and rewatering (six days of irrigation post-drought). Interestingly, drought decreased H2O2 content and activities of superoxide dismutase, catalase (CAT) and ascorbate peroxidase (APX) in the pre-dusk period, although glutathione reductase (GR) and CAM activity increased. Rewatering restored H2O2, but activities of APX, CAT and GR exceeded pre-stress levels in the pre-dusk and/or pre-dawn periods. Results suggest that recovery from a first drought redefines the homeostatic balance of H2O2 scavenging, in which rewatered plants stimulate the enzymatic antioxidant system while drought-exposed plants intensify CAM activity to regulate H2O2 content, a photosynthetic pathway known to prevent oxidative stress. Such data show that young G. monostachia plants adjust CAM and H2O2 scavenging to adapt to water availability.

Proton and anion transport across the tonoplast vesicles in bromeliad species

Crassulacean acid metabolism (CAM) is one of the key innovations in the Neotropical family Bromeliaceae that has enabled many of its species to occupy seasonally water-limited terrestrial environments or microclimatically arid epiphytic niches. However, the relationship between CAM activity and the transport processes responsible for vacuolar organic-acid accumulation at night has not been systematically explored in this family. In the present investigation, ATP- and PPi-dependent proton transport rates were studied in tonoplast membrane vesicles isolated from leaves of six CAM and one C3 species of bromeliads. A consistent feature of these species was the high activity of the tonoplast ATP-driven H+ pump, which, when averaged across the seven species tested, showed a higher specific activity than the tonoplast PPi-driven H+ pump. For all CAM species, the rate of ATP-dependent proton transport into the tonoplast vesicles was strongly influenced by the nature of the balancing organic-acid anion, which displayed the following order of effectiveness: fumarate>malate>citrate. Measurements of leaf organic-acid content in six CAM bromeliads at dusk and dawn showed that nocturnal accumulation of malate exceeded citrate by a factor of ~2.4-20.0-fold in five of six bromeliad species used in this study, demonstrating a close correlation between the CAM rhythm and the intrinsic properties of the vacuolar membrane across which these organic acids are transported.

Implications of leaf ontogeny on drought-induced gradients of CAM expression and ABA levels in rosettes of the epiphytic tank bromeliad Guzmania monostachia

Guzmania monostachia is an epiphytic heteroblastic bromeliad that exhibits rosette leaves forming water-holding tanks at maturity. Different portions along its leaf blades can display variable degrees of crassulacean acid metabolism (CAM) up-regulation under drought. Since abscisic acid (ABA) can act as an important long-distance signal, we conducted a joint investigation of ontogenetic and drought impacts on CAM intensity and ABA levels in different leaf groups within the G. monostachia rosette. For this, three groups of leaves were analysed according to their position within the mature-tank rosette (i.e., younger, intermediate, and older leaves) to characterize the general growth patterns and magnitude of drought-modulated CAM expression. CAM activity was evaluated by analysing key molecules in the biochemical machinery of this photosynthetic pathway, while endogenous ABA content was comparatively measured in different portions of each leaf group after seven days under well-watered (control) or drought treatment. The results revealed that G. monostachia shows more uniform morphological characteristics along the leaves when in the atmospheric stage. The drought treatment of mature-tank rosettes generally induced in older leaves a more severe water loss, followed by the lowest CAM activity and a higher increase in ABA levels, while younger leaves showed an opposite response. Therefore, leaf groups at distinct ontogenetic stages within the tank rosette of G. monostachia responded to drought with variable degrees of water loss and CAM expression. ABA seems to participate in this tissue-compartmented response as a long-distance signalling molecule, transmitting the drought-induced signals originated in older leaves towards the younger ones.

Crassulacean acid metabolism: a continuous or discrete trait?

The key components of crassulacean acid metabolism (CAM) - nocturnal fixation of atmospheric CO2 and its processing via Rubisco in the subsequent light period - are now reasonably well understood in terms of the biochemical reactions defining this water-saving mode of carbon assimilation. Phenotypically, however, the degree to which plants engage in the CAM cycle relative to regular C3 photosynthesis is highly variable. Depending upon species, ontogeny and environment, the contribution of nocturnal CO2 fixation to 24-h carbon gain can range continuously from close to 0% to 100%. Nevertheless, not all possible combinations of light and dark CO2 fixation appear equally common. Large-scale surveys of carbon-isotope ratios typically show a strongly bimodal frequency distribution, with relatively few intermediate values. Recent research has revealed that many species capable of low-level CAM activity are nested within the peak of C3 -type isotope signatures. While questions remain concerning the adaptive significance of dark CO2 fixation in such species, plants with low-level CAM should prove valuable models for investigating the discrete changes in genetic architecture and gene expression that have enabled the evolutionary transition from C3 to CAM.

Influence of germination date on Dioon edule (Zamiaceae) seedling tolerance to water stress

Dioon edule seedling mortality is mostly attributed to dehydration by prolonged drought, even when they present xeromorphic characteristics like the adult plants. The effect of germination date (GD) and soil water deficit on seedling tolerance to water stress was assessed. The seedlings germinated and grown from mature seeds every month from December to April GD were selected to evaluate the leaf area, photosynthetic pigment content, crassulacean acid metabolism (CAM) activity, stomatal conductance (gs) and leaflet anatomy at soil water potential (Ψs) of 0.0MPa (day 1), -0.1MPa (day 40), -1.0MPa (day 90), -1.5MPa (day 130), and a control (0.0MPa at day 130) to recognize differences due to leaf development. The seedlings shifted from C3 to CAM cycling when exposed to water stress at Ψs of -1.0MPa, like adult plants. The March-April GD seedlings with undeveloped sclerified hypodermis and stomata, presented reduced leaf area, lower Chlorophyll a/b ratio, higher CAM activity and midday partial stomatal closure when reached Ψs of -1.0MPa. These have higher probability of dehydration during severe drought (February-April) than those of the December-February GD with similar Ψs. Plants used for restoration purposes must have full leaf development to increase the survival.

Relationships among rainfall, leaf hydrenchyma, and Crassulacean acid metabolism in Pyrrosia lanceolata (L.) Fraw. (Polypodiaceae) in central Taiwan

Leaf succulence is common among drought-adapted plants, including many tropical and subtropical epiphytic species. A prominent anatomical feature of many such succulent leaves is a clear, water-storing tissue often referred to as “hydrenchyma” (water-storage parenchyma). Functionally, hydrenchyma appears to store water for use by the leaf during drought. Although this has been confirmed in several laboratory studies, field studies linking the amount of hydrenchyma in plants with availability of water in their environment are lacking. In this study, the relative amount of leaf hydrenchyma in one of the most widely distributed epiphytes in Taiwan, Pyrrosia lanceolata, was measured in plants growing along a gradient of annual mean precipitation from 2048 to 3688mm. In addition, because Pyrrosia lanceolata is a Crassulacean acid metabolism (CAM) plant, the amount of CAM activity was also examined in plants along the gradient. At each of seven sites along the precipitation gradient, leaves were collected, and, using thin mid-leaf slices, the relative areas of the leaf cross-sections occupied by hydrenchyma were determined. CAM, measured as diel changes in leaf acidity, was measured in plants from each site in the field, after 3 days of water-saturation in the greenhouse, and also after 14 days without water in the greenhouse. Regressions of relative hydrenchyma with ten environmental variables in the dry season revealed that the amount of hydrenchyma was significantly and positively correlated with monthly mean number of rainless days, monthly mean number of days with daily mean temperature over 30°C, and monthly mean temperature. During the wet season, relative hydrenchyma area correlated only with the amount of cloud cover, and the correlation was negative. All plants at all sites exhibited CAM acid fluctuations in the field, under water-saturated conditions, and after desiccation. The largest nocturnal acid accumulations were found when plants were well-hydrated in the field and in the greenhouse, although evidence of drought-induced elevations of CAM was found at the drier sites. The results of this study indicate that the amount of leaf hydrenchyma was greatest in areas with warmer, drier environments. Also, drought-induced elevation of CAM activity occurred in plants from drier sites. This may help to explain the wide range of environments inhabited by this epiphytic fern in Taiwan.

Comparison of photosynthesis and antioxidative protection in Sedum album and Sedum stoloniferum (Crassulaceae) under water stress

Photosynthetic gas exchange, dry mass production, water relations and inducibility of crassulacean acid metabolism (CAM) pathway as well as antioxidative protection during the C3-CAM shift were investigated in Sedum album and Sedum stoloniferum from Crassulaceae under water stress for 20 days. Leaf relative water content (RWC), leaf osmotic and water potential decreased with increasing water stress in both studied species. Significant reduction in dry matter production and leaf thickness was detected only in S. stoloniferum after 20-d water stress. Δtitratable acidity and phosphoenolpyruvate carboxylase (PEPC) activity in S. album responded to drought at early stages of stress treatment, continued to increase throughout the entire stress period and reached levels 15 times higher than those in well-watered plants. In S. stoloniferum, however, both parameters responded later and after a transient increase declined again. In S. stoloniferum, in spite of increase by drought stress, net night-time CO2 assimilation was negative resembling a C3-like pattern of gas exchange. Catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD) activities increased in plants subjected to mild water stress while declined as the stress became severe. Although malondialdehyde (MDA) content was higher in drought-stressed S. stoloniferum, the increase in the concentration of hydrogen peroxide (H2O2) that may act as a signal for C3-CAM transition was higher in S. album compared with S. stoloniferum. In drought-stressed plants, SOD activity showed a clear diurnal fluctuation that was more steadily expressed in S. album. In addition, such pattern was observed for CAT only in S. album. We concluded that temporal and diurnal fluctuation patterns in the activity of antioxidant enzymes depended on duration of drought stress and was related to the mode of photosynthesis and degree of CAM induction. According to our results, S. stoloniferum developed a low degree of CAM activity, e.g. CAM-cycling metabolism, under drought conditions.

Seasonal variation in crassulacean acid metabolism by the aquatic isoetid Littorella uniflora

The seasonal temperature acclimation in crassulacean acid metabolism (CAM) and photosynthetic performance were investigated in the aquatic isoetid, Littorella uniflora. Plants were collected monthly from January to September, and CAM capacity and photosynthesis rates were measured at 5, 10, 15, and 20 °C. Seasonal acclimation was observed for CAM (Q(10) range: 0.6-1.8), and CAM was optimised close to ambient temperature throughout the season. Thus, in winter acclimated L. uniflora, the short-term response to raised temperature resulted in a decline in CAM capacity. Even though the ambient CAM increased from winter to spring/summer, CAM was present in cold acclimated plants, thus indicating an ecophysiological role for CAM even in winter. Similar to CAM, seasonal acclimation was observed in the light and carbon-saturated photosynthesis (Q(10) values ranged from 1.4 to 2.3), and the photosynthetic capacity was generally higher during the winter at all temperatures, indicating compensatory investments in the photosynthetic apparatus. Thus, L. uniflora displayed seasonal temperature acclimation with respect to both CAM and photosynthesis. The estimated in situ contribution of CAM to the carbon budget in L. uniflora was independent of season and varied from 23 to 46 %. A positive correlation between photosynthetic capacity and CAM capacity (both measured in the lab at temperature close to ambient temperature) was found, and the ratio of CAM activity to photosynthetic capacity was higher in summer compared with winter plants. Overall, the results from the present study support the suggested role of CAM as a carbon conserving mechanism of importance for survival in a carbon-limited habitat.

PRE- AND POSTHARVEST METABOLISM OF CROWN LEAVES OF PINEAPPLE FRUIT

Crown burn on exported pineapple fruit causes serious economical losses. In this study, crown burn occurrence was hypothesized to be related to Crassulacean Acid Metabolism (CAM). However, the functioning of this metabolism in the crown leaves remained unexplored. Therefore, pre- and postharvest bio-activity of the crown leaves of pineapple fruit were studied. With the fruit still attached to the plant, CAM activity was detected in the crown leaves. While post-transport diurnal light cycle conditions seemed to reactivate CAM in the crown leaves, no CAM activation was registered during cold-storage transport (simulated) under dark conditions. The substantial organic acid increase in the crown leaves during transport is thought to originate from the pineapple fruit itself and seems to be the major cause of leaf browning.