Determination of the maximum bioaccumulation capacity of various metals in leaves of two Tillandsia species.

Tillandsia species are plants from the Bromeliaceae family which display biomonitoring capacities in both active and passive modes. The bioaccumulation potential of Tillandsia aeranthos (Loisiel.) Desf. and Tillandsia bergeri Mez acclimated to Southern/Mediterranean Europe has never been studied. More generally, few studies have detailed the maximum accumulation potential of Tillandsia leaves through controlled experiments. The aim of this study is to evaluate the maximum accumulation values of seven metals (Co, Cu, Mn, Ni, Pb, Pt, and Zn) in T. aeranthos and T. bergeri leaves. Plants were immersed in different mono elemental metallic solutions of Co (II), Cu (II), Mn (II), Ni (II), Pb (II), Pt (IV), and Zn (II) ions at different concentrations. In addition, cocktail solutions of these seven metals at different concentrations were prepared to study the main differences and the potential selectivity between metals. After exposure, the content of these metals in the leaves were measured by inductively coupled plasma-optical emission spectrometry. Data sets were evaluated by a fitted regression hyperbola model and principal component analysis, maximum metal loading capacity, and thermodynamic affinity constant were determined. The results showed important differences between the two species, with T. bergeri demonstrating higher capacity and affinity for metals than T. aeranthos. Furthermore, between the seven metals, Pb and Ni showed higher enrichment factors (EF). T. bergeri might be a better bioaccumulator than T. aeranthos with marked selectivity for Pb and Ni, metals of concern in air quality biomonitoring.

Radon adsorption and air purification by Spanish moss (Tillandsia usneoides) and its metabolic response to radon exposure.

The capacity of Spanish moss (Tillandsia usneoides), an aerial plant, to adsorb radon (Rn) and absorb CO2 was assessed to analyze its capacity to remove pollutants from indoor air and to determine its radon (Rn) tolerance mechanism. Transcriptomics and metabolomics techniques were used to analyze the response of the plant to Rn exposure. Spanish moss absorbed indoor CO2 at night using the type of photosynthesis termed crassulacean acid metabolism. The CO2 absorption efficiency of the plant was mainly affected by the light duration and diurnal temperature differences. The highest purification efficiency was 48.25%, and the scales on the Spanish moss leaf surface were the key sites for Rn adsorption. Metabolome analysis showed that Rn exposure induced differential metabolites significantly enriched in the metabolism of lipids, amino acids, nucleotides, and carbohydrates. Transcriptome analysis showed significantly upregulated expression levels of functional genes in Rn-exposed leaves. Rn had significant effects on respiratory metabolism, as indicated by upregulated expression of metabolites and functional genes related to the glycolysis pathway, pyruvate oxidation, tricarboxylic acid cycle, and oxidative phosphorylation pathway. These responses indicated that the internal mechanism by which Spanish moss alleviates Rn stress involves an enhancement of cellular energy supplies and regulation of respiratory metabolic pathways to allow adaptation to Rn pollution.

The photosynthetic apparatus of the CAM plant Tillandsia flabellate and its response to water deficit.

CAM plants are superior to C3 plants in drought resistance because of their peculiar photosynthesis pathway and morphological features. While those aspects have been studied for decades, little is known about the photosynthetic machinery of CAM plants. Here, we used a combination of biochemical and biophysical methods to study the photosynthetic apparatus of Tillandsia flabellate, an obligatory CAM plant. Most of the Photosystems super- and sub-complexes have properties very similar to those of Arabidopsis, with the main difference that in Tillandsia PSI-LHCI complexes bind extra LHCI. Functional measurements show that the PSI/PSII ratio is rather low compared to other plants and that the antenna size of both PSI and PSII is small. Upon 30-day water deficiency, the composition of the photosystems does not change significantly, PSII efficiency remains high and no Photosystem II photoinhibition was detected despite a reduction of non-photochemical quenching (NPQ).

Uptake of the natural radioactive gas radon by an epiphytic plant.

Radon (222Rn) is a natural radioactive gas and the major radioactive contributor to human exposure. The present effective ways to control Rn contamination are ventilation and adsorption with activated carbon. Plants are believed to be negligible in reducing airborne Rn. Here, we found epiphytic Tillandsia brachycaulos (Bromeliaceae) was effective in reducing airborne Rn via the leaves. Rn concentrations in the Rn chamber after Tillandsia plant treatments decreased more than those in the natural situation. The specialized foliar trichomes densely covering Tillandsia leaves play a major role in the uptake of Rn because the amplified rough leaf surface area facilitates deposition of Rn progeny particles and the powdery epicuticular wax layer of foliar trichomes uptakes liposoluble Rn. The results provide us a new ecological strategy for Rn contamination control, and movable epiphytic Tillandsia plants can be applied widely in Rn removal systems.

Trade-off between soluble protein production and nutritional storage in Bromeliaceae.

BACKGROUND AND AIMS: Bromeliads are able to occupy some of the most nutrient-poor environments especially because they possess absorptive leaf trichomes, leaves organized in rosettes, distinct photosynthetic pathways [C3, Crassulacean acid metabolism (CAM) or facultative C3-CAM], and may present an epiphytic habit. The more derived features related to these traits are described for the Tillandsioideae subfamily. In this context, the aims of this study were to evaluate how terrestrial predators contribute to the nutrition and performance of bromeliad species, subfamilies and ecophysiological types, whether these species differ in their ecophysiological traits and whether the physiological outcomes are consistent among subfamilies and types (e.g. presence/absence of tank, soil/tank/atmosphere source of nutrients, trichomes/roots access to nutrients). METHODS: Isotopic (15N-enriched predator faeces) and physiological methods (analyses of plant protein, amino acids, growth, leaf mass per area and total N incorporated) in greenhouse experiments were used to investigate the ecophysiological contrasts between Tillandsioideae and Bromelioideae, and among ecophysiological types when a predatory anuran contributes to their nutrition. KEY RESULTS: It was observed that Bromelioideae had higher concentrations of soluble protein and only one species grew more (Ananas bracteatus), while Tillandsioideae showed higher concentrations of total amino acids, asparagine and did not grow. The ecophysiological types that showed similar protein contents also had similar growth. Additionally, an ordination analysis showed that the subfamilies and ecophysiological types were discrepant considering the results of the total nitrogen incorporated from predators, soluble protein and asparagine concentrations, relative growth rate and leaf mass per area. CONCLUSIONS: Bromeliad subfamilies showed a trade-off between two strategies: Tillandsioideae stored nitrogen into amino acids possibly for transamination reactions during nutritional stress and did not grow, whereas Bromelioideae used nitrogen for soluble protein production for immediate utilization, possibly for fast growth. These results highlight that Bromeliaceae evolution may be directly associated with the ability to stock nutrients.

Tillandsia stricta Sol (Bromeliaceae) leaves as monitors of airborne particulate matter-A comparative SEM methods evaluation: Unveiling an accurate and odd HP-SEM method.

Airborne particulate matter (PM) has been included among the most important air pollutants by governmental environment agencies and academy researchers. The use of terrestrial plants for monitoring PM has been widely accepted, particularly when it is coupled with SEM/EDS. Herein, Tillandsia stricta leaves were used as monitors of PM, focusing on a comparative evaluation of Environmental SEM (ESEM) and High-Pressure SEM (HPSEM). In addition, specimens air-dried at formaldehyde atmosphere (AD/FA) were introduced as an SEM procedure. Hydrated specimen observation by ESEM was the best way to get information from T. stricta leaves. If any artifacts were introduced by AD/FA, they were indiscernible from those caused by CPD. Leaf anatomy was always well preserved. PM density was determined on adaxial and abaxial leaf epidermis for each of the SEM proceedings. When compared with ESEM, particle extraction varied from 0 to 20% in air-dried leaves while 23-78% of particles deposited on leaves surfaces were extracted by CPD procedures. ESEM was obviously the best choice over other methods but morphological artifacts increased in function of operation time while HPSEM operation time was without limit. AD/FA avoided the shrinkage observed in the air-dried leaves and particle extraction was low when compared with CPD. Structural and particle density results suggest AD/FA as an important methodological approach to air pollution biomonitoring that can be widely used in all electron microscopy labs. Otherwise, previous PM assessments using terrestrial plants as biomonitors and performed by conventional SEM could have underestimated airborne particulate matter concentration.

The influence of atmospheric particles on the elemental content of vegetables in urban gardens of Sao Paulo, Brazil.

Although urban horticulture provides multiple benefits to society, the extent to which these vegetables are contaminated by the absorption of chemical elements derived from atmospheric deposition is unclear. This study was designed to evaluate the influence of air pollution on leafy vegetables in community gardens of Sao Paulo, Brazil. Vegetable seedlings of Brassica oleracea var. acephala (collard greens) and Spinacia oleracea (spinach) obtained in a non-polluted rural area and growing in vessels containing standard uncontaminated soil were exposed for three consecutive periods of 30, 60 and 90 days in 10 community gardens in Sao Paulo and in one control site. The concentrations of 17 chemical elements (traffic-related elements and those essential to plant biology) were quantified by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Tillandsia usneoides L. specimens were used as air plant biomonitors. The concentrations of As, Cd, Cr and Pb found in vegetables were compared to the recommended values for consumption. Principal Component Analysis (PCA) was used to cluster the elemental concentrations, and Generalized Linear Models (GLMs) were employed to evaluate the association of the factor scores from each PCA component with variables such as local weather, traffic burden and vertical barriers adjacent to the gardens. We found significant differences in the elemental concentrations of the vegetables in the different community gardens. These differences were related to the overall traffic burden, vertical obstacles and local weather. The Pb and Cd concentrations in both vegetables exceeded the limit values for consumption after 60 days of exposure. A strong correlation was observed between the concentration of traffic-related elements in vegetables and in Tillandsia usneoides L. An exposure response was observed between traffic burden and traffic-derived particles absorbed in the vegetables. Traffic-derived air pollution directly influences the absorption of chemical elements in leafy vegetables, and the levels of these elements may exceed the recommended values for consumption.

Suitability of Tillandsia usneoides and Aechmea fasciata for biomonitoring toxic elements under tropical seasonal climate.

Aechmea fasciata was evaluated for the first time as a biomonitor of toxic elements, in comparison to the biomonitoring capacity of Tillandsia usneoides, a well-established biomonitor bromeliad species. Plants of both species were exposed to air pollutants from industrial, urban, and agricultural sources, under the tropical seasonal climate, from June/2011 to April/2013, in five sites of São Paulo State, Brazil, for 8 consecutive exposure periods of 12 weeks each. The levels of essential and non-essential elements, including trace metals, were quantified at the end of each exposure. T. usneoides and A. fasciata indicated N, Fe, Zn, Co, Cr, and V as air contaminants in the studied sites, during wet and dry seasons and both species were recommended for qualitative biomonitoring. Concentration levels of N, Ca, S, Fe, Zn, Cu, B, Co, and Ni were significantly higher in T. usneoides than in A. fasciata. However, A. fasciata showed a higher effective retention capacity of Ni, Pb, V, Cu, Fe, Cr, and Co during field exposure, as indicated by the estimate of enrichment factor relative to basal concentrations. This species is more suitable for detecting the atmospheric pollution level of those metals than the T. usneoides. Both species indicated adequately the seasonal differences in the pollution levels of several elements, but T. usneoides presented higher ability for biomonitoring the spatial variations and for indicating more properly the sources of each element in the studied region than the A. fasciata.

Bioindicating potential of strontium contamination with Spanish moss Tillandsia usneoides.

Tillandsia species have been recognized as efficient biomonitors of air pollution, but rarely exploited in bioindicating of strontium, an important nuclide. We exposed Tillandsia usneoides, colloquially known as Spanish moss due to its filamentous morphology but is an atypical angiosperm in the family Bromeliaceae, to the solutions with different Sr concentrations (0.1-100 mmol/L). The results showed that plants were able to endure Sr stress for a relatively long period, which suggests that T. usneoides is able to resist this toxic element. T. usneoides had the highest uptake ratio of Sr (82.21 ± 0.12%) when the plants were exposed to 0.1 mmol/L Sr solutions. Sr contents in T. usneoides increased significantly with the increase in applied metal solution concentrations. Low Sr stimulated the formation of chlorophyll, but high Sr decreased the contents of chlorophyll, and no significant effect on the total biomass was found in T. usneoides. In contrast, the permeability of plasma membrane based on the relative electronic conductivity in T. usneoides increased significantly under Sr stress, indicating that Sr probably caused oxidative stress. Moreover, correlation analysis showed that the leaf relative conductivity was significantly positively correlated with Sr contents in the plants after Sr treatments. Therefore, T. usneoides has considerable potential for monitoring Sr polluted environments through measuring Sr contents in the plant directly or exploiting the leaf relative conductivity as an indirect biomarker.

Tillandsia usneoides: a successful alternative for biomonitoring changes in air quality due to a new highway in São Paulo, Brazil.

Tillandsia usneoides is an aerial epiphytic bromeliad that absorbs water and nutrients directly from the atmosphere by scales covering its surface. We expanded the use of this species as a broader biomonitor based on chemical and structural markers to detect changes in air quality. The usefulness of such comprehensive approach was tested during the construction and opening of a highway (SP-21) in São Paulo State, Brazil. The biomonitoring study was performed from 2009 to 2012, thus comprising the period during construction and after the highway inauguration. Metal accumulation and structural alterations were assessed, in addition to microscopy analyses to understand the metal chelation in plant tissues and to assess the causes of alterations in the number and shape of scale cells. Altogether, our analyses support the use of this species as a wide biomonitor of air quality in urbanized areas.

Foliar trichome-aided formaldehyde uptake in the epiphytic Tillandsia velutina and its response to formaldehyde pollution.

Epiphytic Tillandsia (Bromeliaceae) species have been found to be efficient biomonitors of atmospheric heavy metals and persistent organic pollutants, but have not been used to monitor or remove the primary indoor atmospheric pollutant formaldehyde (FA). The absorptive capacity of Tillandsia trichomes is well-established, but potential secondary effects of foliar trichomes on gas exchange remain unclear. Our study investigated whether Tillandsia species can absorb FA efficiently and if the leaf trichomes function to improve FA uptake, using Tillandsia velutina. Plants with intact trichomes, decreased FA concentration by 48.42% in 12 h from 1060 µg m(-3) to 546.67 µg m(-3), while FA concentration decreased only by 22.51% in the plants without trichomes. Moreover, the more trichomes removed from the leaves, the lower the capability of FA uptake per unit leaf area, which suggested that T. velutina was capable of absorbing a large amount of FA via the leaves and specialized trichomes facilitated the whole leaf tissue FA absorption. In addition, all plants exposed to FA were chloric, had a reduction in measured leaf chlorophyll, and an increment in permeability of plasma membranes. However, plants in which trichomes had been removed declined or increased more quickly than plants with intact trichomes, indicating Tillandsia leaf trichomes also give the leaves some protection against this toxin.

Comparison of oxidative stress in four Tillandsia species exposed to cadmium.

This is first study comparing four morphologically variable species of the genus Tillandsia and therefore various responses to the cadmium (Cd) action were expected. In accordance, Cd accumulation increased in order Tillandsia fasciculata < Tillandsia brachycaulos < Tillandsia pruinosa < Tillandsia capillaris, reaching 29.6 and 197.4 µg g(-1) DW in first and last species after watering with 2 µM Cd(2+) solution over 30 days. Fluorescence visualization of oxidative stress confirmed increase in ROS and especially elevation in hydroperoxides though no visible symptoms appeared on the plants. At the same time, nitric oxide generation and nitroso-glutathione depletion by Cd treatment were typically observed. Fluorescence staining of Cd using two dyes (PhenGreen and Leadmium) showed that Leadmium fits better with AAS quantification. Macro- and micro-nutrients were not considerably affected except for zinc. Reduced glutathione content was the highest in control T. fasciculata while oxidized glutathione in T. capillaris. Ascorbic acid amount revealed extreme quantitative differences among species and decreased in T. fasciculata only. Free amino acids accumulation was similar among species except for T. capillaris and Cd caused both depletion and increase but without high quantitative differences. Data are explanatively discussed in the context of limited literature related to oxidative stress in epiphytic plants and with general responses of plants to cadmium/heavy metals.

Reconstructing the δ(18) O of atmospheric water vapour via the CAM epiphyte Tillandsia usneoides: seasonal controls on δ(18) O in the field and large-scale reconstruction of δ(18) Oa.

Using both oxygen isotope ratios of leaf water (δ(18) OL ) and cellulose (δ(18) OC ) of Tillandsia usneoides in situ, this paper examined how short- and long-term responses to environmental variation and model parameterization affected the reconstruction of the atmospheric water vapour (δ(18) Oa ). During sample-intensive field campaigns, predictions of δ(18) OL matched observations well using a non-steady-state model, but the model required data-rich parameterization. Predictions from the more easily parameterized maximum enrichment model (δ(18) OL-M ) matched observed δ(18) OL and observed δ(18) Oa when leaf water turnover was less than 3.5 d. Using the δ(18) OL-M model and weekly samples of δ(18) OL across two growing seasons in Florida, USA, reconstructed δ(18) Oa was -12.6 ± 0.3‰. This is compared with δ(18) Oa of -12.4 ± 0.2‰ resolved from the growing-season-weighted δ(18) OC . Both of these values were similar to δ(18) Oa in equilibrium with precipitation, -12.9‰. δ(18) Oa was also reconstructed through a large-scale transect with δ(18) OL and the growing-season-integrated δ(18) OC across the southeastern United States. There was considerable large-scale variation, but there was regional, weather-induced coherence in δ(18) Oa when using δ(18) OL . The reconstruction of δ(18) Oa with δ(18) OC generally supported the assumption of δ(18) Oa being in equilibrium with precipitation δ(18) O (δ(18) Oppt ), but the pool of δ(18) Oppt with which δ(18) Oa was in equilibrium - growing season versus annual δ(18) Oppt - changed with latitude.

Air pollutant characterization in Tula industrial corridor, Central Mexico, during the MILAGRO study.

Pollutant emissions and their contribution to local and regional air quality at the industrial area of Tula were studied during a four-week period as part of the MILAGRO initiative. A recurrent shallow stable layer was observed in the morning favoring air pollutants accumulation in the lower 100 m atmospheric layer. In the afternoon the mixing layer height reached 3000 m, along with a featuring low level jet which was responsible of transporting air pollutants at regional scales. Average PM10 at Jasso (JAS) and Tepeji (TEP) was 75.1 and 36.8 µ g/m(3), respectively while average PM2.5 was 31.0 and 25.7 µ g/m(3). JAS was highly impacted by local limestone dust, while TEP was a receptor of major sources of combustion emissions with 70% of the PM10 constituted by PM2.5. Average hourly aerosol light absorption was 22 Mm(-1), while aerosol scattering (76 Mm(-1)) was higher compared to a rural site but much lower than at Mexico City. δ(13)C values in the epiphyte Tillandsia recurvata show that the emission plume directly affects the SW sector of Mezquital Valley and is then constrained by a mountain range preventing its dispersion. Air pollutants may exacerbate acute and chronic adverse health effects in this region.

Potential of monitoring nuclides with the epiphyte Tillandsia usneoides: Uptake and localization of 133Cs.

Epiphytic Tillandsia plants are efficient air pollution biomonitors and traditionally used to monitor atmospheric heavy metal pollution, but rarely nuclides monitoring. Here we evaluated the potential of Tillandsia usneoides for monitoring (133)Cs and investigated if Cs was trapped by the plant external surface structures. The results showed that T. usneoides was able to survive relatively high Cs stress. With the increase of Cs solution concentration, the total of Cs in plants increased significantly, which suggests that the plants could accumulate Cs quickly and effectively. Therefore, T. usneoides has considerable potential for monitoring Cs polluted environments. In addition, scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS) analysis showed that Cs was detected in each type of cells in foliar trichomes, and the ratio of Cs in the internal disc cell was higher than that in ring cell and wing cell, which indicates that the mechanism of adsorption Cs in Tillandsia has an active component.

Physiological responses of Tillandsia albida (Bromeliaceae) to long-term foliar metal application.

The impact of 2-month foliar application of cadmium, nickel and their combination (10 µM) on Tillandsia albida was studied. Cadmium caused damage of tissue but assimilation pigments were depressed in Cd+Ni variant only. Stress-related parameters (ROS and peroxidase activities) were elevated by Cd and Cd+Ni while MDA content remained unaffected. Free amino acids accumulated the most in Ni alone but soluble proteins were not influenced. Among phenolic acids, mainly vanillin contributed to increase of their sum in all variants while soluble phenols even decreased in Cd+Ni and flavonols slightly increased in Cd variants. Phenolic enzymes showed negligible responses to almost all treatments. Mineral nutrients (K, Ca, Na, Mg, Fe, and Zn) were not affected by metal application but N content increased. Total Cd or Ni amounts reached over 400 µg g(-1) DW and were not affected if metal alone and combined treatment is compared while absorbed content differed (ca. 50% of total Cd was absorbed while almost all Ni was absorbed). These data indicate tolerance of T. albida to foliar metal application and together with strong xerophytic morphology, use for environmental studies is recommended.

On the controls of leaf-water oxygen isotope ratios in the atmospheric Crassulacean acid metabolism epiphyte Tillandsia usneoides.

Previous theoretical work showed that leaf-water isotope ratio (δ(18)O(L)) of Crassulacean acid metabolism epiphytes was controlled by the δ(18)O of atmospheric water vapor (δ(18)O(a)), and observed δ(18)O(L) could be explained by both a non-steady-state model and a "maximum enrichment" steady-state model (δ(18)O(L-M)), the latter requiring only δ(18)O(a) and relative humidity (h) as inputs. δ(18)O(L), therefore, should contain an extractable record of δ(18)O(a). Previous empirical work supported this hypothesis but raised many questions. How does changing δ(18)O(a) and h affect δ(18)O(L)? Do hygroscopic trichomes affect observed δ(18)O(L)? Are observations of changes in water content required for the prediction of δ(18)O(L)? Does the leaf need to be at full isotopic steady state for observed δ(18)O(L) to equal δ(18)O(L-M)? These questions were examined with a climate-controlled experimental system capable of holding δ(18)O(a) constant for several weeks. Water adsorbed to trichomes required a correction ranging from 0.5‰ to 1‰. δ(18)O(L) could be predicted using constant values of water content and even total conductance. Tissue rehydration caused a transitory change in δ(18)O(L), but the consequent increase in total conductance led to a tighter coupling with δ(18)O(a). The non-steady-state leaf water models explained observed δ(18)O(L) (y = 0.93*x - 0.07; r(2) = 0.98) over a wide range of δ(18)O(a) and h. Predictions of δ(18)O(L-M) agreed with observations of δ(18)O(L) (y = 0.87*x - 0.99; r(2) = 0.92), and when h > 0.9, the leaf did not need to be at isotopic steady state for the δ(18)O(L-M) model to predict δ(18)O(L) in the Crassulacean acid metabolism epiphyte Tillandsia usneoides.

Assessment of heavy metals in the particulate matter of two Brazilian metropolitan areas by using Tillandsia usneoides as atmospheric biomonitor.

PURPOSE: The aims of this paper were to quantify the heavy metals (HM) in the air of different sites in Rio de Janeiro (RJ) and Salvador (SA) using Tillandsia usneoides (Bromeliaceae) as a biomonitor, and to study the morphology and elemental composition of the air particulate matter (PM) retained on the Tillandsia surface. METHODS: Tillandsia samples were collected in a noncontaminated area and exposed to the air of five sites in RJ State and seven in SA for 45 days, in two seasons. Samples were prepared to HM quantification by flame atomic absorption spectrophotometry, while morphological and elemental characterizations were studied by using scanning electron microscopy. RESULTS: HM concentrations were significantly higher when compared to control sites. We found an increasing metal concentration as follows: Cd < Cr < Pb < Cu < Zn. PM exhibited a morphology varying from amorphous- to polygonal-shaped particles. Size measurements indicated that more than 80% of particles were less than 10 µm. PM contained aluminosilicates iron-rich particles, but Zn, Cu, Cr, and Ba were also detected. CONCLUSION: HM input in the atmosphere was mainly associated with anthropogenic sources such as vehicle exhaust. Elemental analysis detected HM in the inhalable particles, indicating that those HMs may intensify the toxic effects of PM on human health. Our results indicated T. usneoides as an adequate biomonitor of HM in the PM belonging to the inhalable fraction.

Correlation between citric acid and nitrate metabolisms during CAM cycle in the atmospheric bromeliad Tillandsia pohliana.

Crassulacean acid metabolism (CAM) confers crucial adaptations for plants living under frequent environmental stresses. A wide metabolic plasticity can be found among CAM species regarding the type of storage carbohydrate, organic acid accumulated at night and decarboxylating system. Consequently, many aspects of the CAM pathway control are still elusive while the impact of this photosynthetic adaptation on nitrogen metabolism has remained largely unexplored. In this study, we investigated a possible link between the CAM cycle and the nitrogen assimilation in the atmospheric bromeliad Tillandsia pohliana by simultaneously characterizing the diel changes in key enzyme activities and metabolite levels of both organic acid and nitrate metabolisms. The results revealed that T. pohliana performed a typical CAM cycle in which phosphoenolpyruvate carboxylase and phosphoenolpyruvate carboxykinase phosphorylation seemed to play a crucial role to avoid futile cycles of carboxylation and decarboxylation. Unlike all other bromeliads previously investigated, almost equimolar concentrations of malate and citrate were accumulated at night. Moreover, a marked nocturnal depletion in the starch reservoirs and an atypical pattern of nitrate reduction restricted to the nighttime were also observed. Since reduction and assimilation of nitrate requires a massive supply of reducing power and energy and considering that T. pohliana lives overexposed to the sunlight, we hypothesize that citrate decarboxylation might be an accessory mechanism to increase internal CO2 concentration during the day while its biosynthesis could provide NADH and ATP for nocturnal assimilation of nitrate. Therefore, besides delivering photoprotection during the day, citrate might represent a key component connecting both CAM pathway and nitrogen metabolism in T. pohliana; a scenario that certainly deserves further study not only in this species but also in other CAM plants that nocturnally accumulate citrate.

Carbon stable isotopic composition of soluble sugars in Tillandsia epiphytes varies in response to shifts in habitat.

We studied C stable isotopic composition (delta(13)C) of bulk leaf tissue and extracted sugars of four epiphytic Tillandsia species to investigate flexibility in the use of crassulacean acid metabolism (CAM) and C(3) photosynthetic pathways. Plants growing in two seasonally dry tropical forest reserves in Mexico that differ in annual precipitation were measured during wet and dry seasons, and among secondary, mature, and wetland forest types within each site. Dry season sugars were more enriched in (13)C than wet season sugars, but there was no seasonal difference in bulk tissues. Bulk tissue delta(13)C differed by species and by forest type, with values from open-canopied wetlands more enriched in (13)C than mature or secondary forest types. The shifts within forest habitat were related to temporal and spatial changes in vapor pressure deficits (VPD). Modeling results estimate a possible 4% increase in the proportional contribution of the C(3) pathway during the wet season, emphasizing that any seasonal or habitat-mediated variation in photosynthetic pathway appears to be quite moderate and within the range of isotopic effects caused by variation in stomatal conductance during assimilation through the C(3) pathway and environmental variation in VPD. C isotopic analysis of sugars together with bulk leaf tissue offers a useful approach for incorporating short- and long-term measurements of C isotope discrimination during photosynthesis.