RESUMO
The common ice plant (Mesembryanthemum crystallinum L.) is a facultative crassulacean acid metabolism (CAM) plant, and its ability to recover from stress-induced CAM has been confirmed. We analysed the photosynthetic metabolism of this plant during the 72-h response period following salinity stress removal from three perspectives. In plants under salinity stress (CAM) we found a decline of the quantum efficiencies of PSII (Y(II)) and PSI (Y(I)) by 17% and 15%, respectively, and an increase in nonphotochemical quenching (NPQ) by almost 25% in comparison to untreated control. However, 48 h after salinity stress removal, the PSII and PSI efficiencies, specifically Y(II) and Y(I), elevated nonphotochemical quenching (NPQ) and donor side limitation of PSI (YND), were restored to the level observed in control (C3 plants). Swelling of the thylakoid membranes, as well as changes in starch grain quantity and size, have been found to be components of the salinity stress response in CAM plants. Salinity stress induced an over 3-fold increase in average starch area and over 50% decline of average seed number in comparison to untreated control. However, in plants withdrawn from salinity stress, during the first 24 h of recovery, we observed chloroplast ultrastructures closely resembling those found in intact (control) ice plants. Rapid changes in photosystem functionality and chloroplast ultrastructure were accompanied by the induction of the expression (within 24 h) of structural genes related to the PSI and PSII reaction centres, including PSAA, PSAB, PSBA (D1), PSBD (D2) and cp43. Our findings describe one of the most flexible photosynthetic metabolic pathways among facultative CAM plants and reveal the extent of the plasticity of the photosynthetic metabolism and related structures in the common ice plant.
Assuntos
Metabolismo Ácido das Crassuláceas/genética , Mesembryanthemum/genética , Fotossíntese/genética , Estresse Salino/genética , Cloroplastos/efeitos dos fármacos , Cloroplastos/genética , Metabolismo Ácido das Crassuláceas/efeitos dos fármacos , Mesembryanthemum/efeitos dos fármacos , Fotossíntese/efeitos dos fármacos , Plastídeos/efeitos dos fármacos , Plastídeos/genética , Salinidade , Estresse Salino/efeitos dos fármacos , Cloreto de Sódio/farmacologia , Amido/genética , Tilacoides/efeitos dos fármacos , Tilacoides/genéticaRESUMO
Mesembryanthemum crystallinum (common ice plant) is a halophyte species that has adapted to extreme conditions. In this study, we cloned a McHB7 transcription factor gene from the ice plant. The expression of McHB7 was significantly induced by 500 mM NaCl and it reached the peak under salt treatment for 7 days. The McHB7 protein was targeted to the nucleus. McHB7-overexpressing in ice plant leaves through Agrobacterium-mediated transformation led to 25 times more McHB7 transcripts than the non-transformed wild type (WT). After 500 mM NaCl treatment for 7 days, the activities of superoxide dismutase (SOD) and peroxidase (POD) and water content of the transgenic plants were higher than the WT, while malondialdehyde (MDA) was decreased in the transgenic plants. A total of 1082 and 1072 proteins were profiled by proteomics under control and salt treatment, respectively, with 22 and 11 proteins uniquely identified under control and salt stress, respectively. Among the 11 proteins, 7 were increased and 4 were decreased after salt treatment. Most of the proteins whose expression increased in the McHB7 overexpression (OE) ice plants under high salinity were involved in transport regulation, catalytic activities, biosynthesis of secondary metabolites, and response to stimulus. The results demonstrate that the McHB7 transcription factor plays a positive role in improving plant salt tolerance.
Assuntos
Mesembryanthemum/metabolismo , Proteínas de Plantas/metabolismo , Proteômica , Tolerância ao Sal/fisiologia , Sequência de Aminoácidos , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Biologia Computacional , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ontologia Genética , Mesembryanthemum/efeitos dos fármacos , Mesembryanthemum/genética , Filogenia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Transporte Proteico/efeitos dos fármacos , Salinidade , Tolerância ao Sal/efeitos dos fármacos , Tolerância ao Sal/genética , Cloreto de Sódio/farmacologia , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética , Frações Subcelulares/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Many areas exhibiting increased concentrations of soluble salts are simultaneously polluted with heavy metals (HM), and halophytes with extended tolerance to heavy metal toxicity seem to represent a promising tool for their phytoremediation. In this study, the response of the soil-grown C3-CAM (Crassulacean acid metabolism) intermediate halophyte Mesembryanthemum crystallinum (common ice plant) to increased concentrations of Cd (0.01-1â¯mM) was investigated. None of the tested Cd treatments affected growth parameters or tissue water content of either C3 or CAM-performing plants. Chlorophyll a fluorescence confirmed high tolerance of the photosynthetic apparatus of both metabolic states towards Cd. Plants performing both photosynthesis types accumulated significant Cd amounts only under the highest (1â¯mM) treatment, and the metal was primarily deposited in the roots, which are features typical of an excluding strategy. Upon the application of 1â¯mM Cd solution CAM-performing plants, due to the NaCl pre-treatment applied for CAM induction, were exposed to significantly higher amounts of bioavailable Cd in comparison with those of C3-performing plants. As a result, roots of CAM plants accumulated over 4-fold higher Cd amounts when compared with C3 plants. In our opinion, enhanced Cd-accumulating potential observed in CAM-performing plants was the effect of osmotic stress episode and resulting modifications e.g. in the detoxifying capacity of the antioxidative system. Increased antioxidative potential of NaCl pre-treated plants was pronounced with significantly higher activity of CuZnSOD (copper-zinc superoxide dismutase), not achievable in C3 plants subjected to high Cd concentrations. Moreover, the applied Cd doses induced SOD activity in a compartment-dependent manner only in C3 plants. We confirmed that none of the applied Cd concentrations initiated the metabolic shift from C3 to CAM.
Assuntos
Cádmio/efeitos adversos , Mesembryanthemum/efeitos dos fármacos , Plantas Tolerantes a Sal/efeitos dos fármacos , Poluentes do Solo/efeitos adversos , Relação Dose-Resposta a Droga , Mesembryanthemum/enzimologia , Mesembryanthemum/crescimento & desenvolvimento , Mesembryanthemum/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/enzimologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/enzimologia , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Plantas Tolerantes a Sal/enzimologia , Plantas Tolerantes a Sal/crescimento & desenvolvimento , Plantas Tolerantes a Sal/metabolismo , Superóxido Dismutase/metabolismoRESUMO
Exogenously applied H2O2 (50, 100 and 200mM) to Mesembryanthemum crystallinum root medium induced a transition from C3 to Crassulacean Acid Metabolism (CAM), as evaluated by diurnal malate (Δmal) fluctuations. A very high concentration of H2O2 (400mM) reduced Δmal below the value measured in control plants. An increase of malate content during the night in 400mM H2O2-treated plants might suggest that malate decarboxylation is crucial for CAM functioning. We conclude that malate plays a dual role: i) a protective and signaling function before CAM expression, and ii) a storage form of CO2 in plants performing CAM. A slight stimulation of photosystem II (PSII) photochemistry and net photosynthesis observed during the C3-CAM shift indicated that neither photoinhibition nor reduction of the photosynthetic rate were prerequisites for CAM. Moreover, CAM induction corresponded to a decrease of catalase activity. In CAM-performing plants, α-tocopherol, polyamines (putrescine and spermidine) and proline showed daily alterations and the content of α-tocopherol and polyamines was lower at the end of the day. In contrast, the proline concentration correlated with the applied H2O2 concentration and was higher at the end of the day in treated plants. The dynamic changes of antioxidant and osmolyte levels suggest their active role in preventing oxidative damage, stress acclimation mechanisms and involvement in metabolic regulation and/or signal transduction cascades.
Assuntos
Antioxidantes/metabolismo , Carbono/metabolismo , Ácidos Carboxílicos/metabolismo , Peróxido de Hidrogênio/farmacologia , Mesembryanthemum/fisiologia , Osmose/efeitos dos fármacos , Plantas Tolerantes a Sal/fisiologia , Catalase/metabolismo , Gases/metabolismo , Mesembryanthemum/efeitos dos fármacos , Mesembryanthemum/enzimologia , Fotossíntese/efeitos dos fármacos , Complexo de Proteína do Fotossistema II/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Poliaminas/metabolismo , Prolina/metabolismo , Plantas Tolerantes a Sal/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacos , alfa-Tocoferol/metabolismoRESUMO
Mesembryanthemum crystallinum (ice plant) exhibits extreme tolerance to salt. Epidermal bladder cells (EBCs), developing on the surface of aerial tissues and specialized in sodium sequestration and other protective functions, are critical for the plant's stress adaptation. We present the first transcriptome analysis of EBCs isolated from intact plants, to investigate cell type-specific responses during plant salt adaptation. We developed a de novo assembled, nonredundant EBC reference transcriptome. Using RNAseq, we compared the expression patterns of the EBC-specific transcriptome between control and salt-treated plants. The EBC reference transcriptome consists of 37 341 transcript-contigs, of which 7% showed significantly different expression between salt-treated and control samples. We identified significant changes in ion transport, metabolism related to energy generation and osmolyte accumulation, stress signalling, and organelle functions, as well as a number of lineage-specific genes of unknown function, in response to salt treatment. The salinity-induced EBC transcriptome includes active transcript clusters, refuting the view of EBCs as passive storage compartments in the whole-plant stress response. EBC transcriptomes, differing from those of whole plants or leaf tissue, exemplify the importance of cell type-specific resolution in understanding stress adaptive mechanisms.
Assuntos
Mesembryanthemum/citologia , Mesembryanthemum/genética , Epiderme Vegetal/citologia , Epiderme Vegetal/genética , Salinidade , Transcriptoma/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ontologia Genética , Redes Reguladoras de Genes/efeitos dos fármacos , Mesembryanthemum/efeitos dos fármacos , Anotação de Sequência Molecular , Epiderme Vegetal/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Cloreto de Sódio/farmacologia , Estresse Fisiológico/efeitos dos fármacos , Transcriptoma/efeitos dos fármacosRESUMO
The effect of abscisic acid (ABA) and fluridone on the content of endogenous phytohormones and free polyamines and the intensity of oxidative stress was studied in plants of Mesembryanthemum crystallinum L. under salinity. It was shown that the pretreatment of plant roots with 1 µM ABA, followed by the action of 300 mM NaCl, caused a protective effect and improved the physiological state of the plants, which was manifested in increased biomass and content of available cytokinins and reduced values of the indicators of oxidative stress. It was noted that the inhibitor fluridone reduced the effect of ABA and acted as a pro-oxidant.
Assuntos
Ácido Abscísico/farmacologia , Mesembryanthemum/efeitos dos fármacos , Reguladores de Crescimento de Plantas/metabolismo , Poliaminas/metabolismo , Piridonas/farmacologia , Ácido Abscísico/metabolismo , Citocininas/metabolismo , Mesembryanthemum/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Prolina/metabolismo , Salinidade , Cloreto de Sódio/farmacologiaRESUMO
KEY MESSAGE: H2O2 is necessary to elicit rhizogenic action of auxin. Activities of specific catalase and manganese superoxide dismutase forms mark roots development. Hypocotyl explants of Mesembryanthemum crystallinum regenerated roots on medium containing 2,4-dichlorophenoxyacetic acid. Explants became competent to respond to the rhizogenic action of auxin on day 3 of culture, when hydrogen peroxide content in cultured tissue was the highest. L-Ascorbic acid added to the medium at 5 µM lowered the H2O2 level, inhibited rhizogenesis and induced non-regenerative callus, suggesting that certain level of H2O2 is required to promote root initiation. Coincident with the onset of rhizogenic determination, meristemoids formed at the periphery of the hypocotyl stele and the activity of the manganese form of superoxide dismutase, MnSOD-2 was induced. Once induced, MnSOD-2 activity was maintained through the post-determination phase of rooting, involving root growth. MnSOD-2 activity was not found in non-rhizogenic explants maintained in the presence of AA. Analyses of the maximum photochemical efficiency of photosystem II and the oxygen uptake rate revealed that the explants were metabolically arrested during the predetermination stage of rhizogenesis. Respiratory and photosynthetic rates were high during root elongation and maturation. Changes in catalase and peroxidase activities correlated with fluctuations of endogenous H2O2 content throughout rhizogenic culture. Expression of a specific CAT-2 form accompanied the post-determination stage of rooting and a high rate of carbohydrate metabolism during root growth. On the other hand, the occurrence of MnSOD-2 activity did not depend on the metabolic status of explants. The expression of MnSOD-2 activity throughout root development seems to relate it specifically to root metabolism and indicates it as a molecular marker of rhizogenesis in M. crystallinum.
Assuntos
Antioxidantes/metabolismo , Peróxido de Hidrogênio/metabolismo , Hipocótilo/crescimento & desenvolvimento , Mesembryanthemum/enzimologia , Mesembryanthemum/crescimento & desenvolvimento , Raízes de Plantas/enzimologia , Raízes de Plantas/crescimento & desenvolvimento , Ácido 2,4-Diclorofenoxiacético , Ácido Ascórbico/farmacologia , Catalase/metabolismo , Meios de Cultura/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Guaiacol/farmacologia , Hipocótilo/efeitos dos fármacos , Meristema/efeitos dos fármacos , Meristema/crescimento & desenvolvimento , Mesembryanthemum/efeitos dos fármacos , Oxigênio/metabolismo , Peroxidase/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Raízes de Plantas/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Superóxido Dismutase/metabolismoRESUMO
Mesembryanthemum edule L. is an edible and medicinal halophyte widespread in Tunisia seashore. In this study, parameters of oxidative stress, phenolic compounds and antioxidant activities were comparatively investigated in two M. edule provenances (Jerba and Bizerte, respectively sampled from arid and humid bioclimatic stages). Plants were subjected to 0, 300 and 600mM NaCl treatment under glasshouse conditions. Results showed that M. edule response to salinity depends on provenance (P), salt treatment (T) and their interaction (P×T). (T) affected more significantly the oxidative stress parameters and antioxidant activities than (P) and (P×T). Conversely, (P) was much affluent for tannin polymerization degree and interaction between the two factors (P×T) was more determinants for analyzed antioxidant parameters. The higher salt tolerance of Jerba plants was associated with low levels of malondialdehyde and of electrolyte leakage mainly at 600mM NaCl. Besides, antioxidant activities of Jerba provenance, were more efficient than Bizerte. In addition, avicularin was the major phenolic in both provenances. This compound concentration increased with salinity in Jerba shoots, while it was reduced in Bizerte especially at 600mM NaCl. Overall, the higher salt tolerance of plants from Jerba provenance, and to a lower extent of those from Bizerte, may be partly related to their better capacity to limit oxidative damage when salt-challenged, and this is likely the result of redistribution in phenolic composition. Besides, abiotic factors such as salinity could be determinant in antioxidant potentiality of this medicinal plant.
Assuntos
Antioxidantes/metabolismo , Mesembryanthemum/efeitos dos fármacos , Mesembryanthemum/fisiologia , Fenóis/metabolismo , Cloreto de Sódio/farmacologia , Antioxidantes/análise , Sequestradores de Radicais Livres/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Mesembryanthemum/crescimento & desenvolvimento , Estresse Oxidativo/efeitos dos fármacos , Fenóis/análise , Fenóis/isolamento & purificação , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/fisiologia , Polimerização/efeitos dos fármacos , Tolerância ao Sal , Plantas Tolerantes a SalRESUMO
The effect of free cadaverine (Cad) on its conjugates formation was analyzed in roots of the common ice plants (Mesembryanthemum crystallinum L.). It was found for the first time that Cad could induce oxidative burst in the roots of adult plants, as was evident from the sharp decrease in the content of Cad soluble or insoluble conjugates. This unusual effect was associated with the increased oxidative degradation of exogenous Cad (1mM, 1.5h) and intense H(2)O(2) production in the root cells of adult plants. Root treatment of both juvenile and adult plants with H(2)O(2) (1mM, 1.5h) reduced the content of soluble Cad conjugates and increased the content of their components, free Cad and phenols. We also found that one of the possible reasons of the negative effect of exogenous diamine on the formation of conjugated forms in adult roots was alkalization of the root apoplast at Cad addition to nutrient medium and the unusual O(2)(-) synthase function as a pH-dependent guaiacol peroxidase in the presence of a high content of H(2)O(2). This was confirmed by the data on the accumulation of O(2)(-) and enhanced superoxide dismutase activity in adult roots under treatment with Cad. It is possible that the accumulation of O(2)(-) together with H(2)O(2) was also responsible for oxidative burst, which induced a decrease in the content of Cad conjugates in adult roots of the common ice plants.
Assuntos
Cadaverina/metabolismo , Cadaverina/farmacologia , Mesembryanthemum/citologia , Mesembryanthemum/efeitos dos fármacos , Explosão Respiratória/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/farmacologia , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Mesembryanthemum/enzimologia , NADPH Oxidases/antagonistas & inibidores , Oniocompostos/farmacologia , Peroxidase/metabolismo , Fenóis/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/enzimologia , Cloreto de Sódio/farmacologia , Solubilidade/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacos , Superóxidos/metabolismoRESUMO
The common ice plant (Mesembryanthemum crystallinum L.) has emerged as a useful model for molecular genetic studies of Crassulacean acid metabolism (CAM) because CAM can be induced in this species by water deficit or salinity stress. Non-redundant sequence information from expressed sequence tag data was used to fabricate a custom oligonucleotide microarray to compare large-scale mRNA expression patterns in M. crystallinum plants conducting C(3) photosynthesis versus CAM. Samples were collected every 4 h over a 24 h time period at the start of the subjective second day from plants grown under constant light and temperature conditions in order to capture variation in mRNA expression due to salinity stress and circadian clock control. Of 8455 genes, a total of 2343 genes (approximately 28%) showed a significant change as judged by analysis of variance (ANOVA) in steady-state mRNA abundance at one or more time points over the 24 h period. Of these, 858 (10%) and 599 (7%) exhibited a greater than two-fold ratio (TFR) increase or decrease in mRNA abundance, respectively. Functional categorization of these TFR genes revealed that many genes encoding products that function in CAM-related C(4) acid carboxylation/decarboxylation, glycolysis/gluconeogenesis, polysaccharide, polyol, and starch biosynthesis/degradation, protein degradation, transcriptional activation, signalling, stress response, and transport facilitation, and novel, unclassified proteins exhibited stress-induced increases in mRNA abundance. In contrast, salt stress resulted in a significant decrease in transcript abundance for genes encoding photosynthetic functions, protein synthesis, and cellular biogenesis functions. Many genes with CAM-related functions exhibited phase shifts in their putative circadian expression patterns following CAM induction. This report establishes an extensive catalogue of gene expression patterns for future investigations aimed at understanding the complex, transcriptional hierarchies that govern CAM-specific expression patterns. A novel graph-theoretic approach called 'Max Clique Builder' is introduced that identifies and organizes sets of coordinately regulated genes, such as those encoding subunits of the vacuolar H(+)-ATPase complex, into tighter functionally related clusters with more similar expression patterns compared with standard hierarchical clustering methods.
Assuntos
Perfilação da Expressão Gênica , Mesembryanthemum/metabolismo , Fotossíntese/fisiologia , RNA Mensageiro/metabolismo , Ritmo Circadiano , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Temperatura Alta , Mesembryanthemum/efeitos dos fármacos , Mesembryanthemum/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , RNA de Plantas/metabolismo , Cloreto de Sódio/farmacologiaRESUMO
Mesembryanthemum crystallinum L. (Aizoaceae) is a facultative annual halophyte and a C(3)-photosynthesis/crassulacean acid metabolism intermediate species currently used as a model plant in stress physiology. Both salinity and high light irradiance stress are known to induce CAM in this species. The present study was performed to provide a diagnosis of alterations at the photosystem II level during salinity and irradiance stress. Plants were subjected for up to 13 days to either 0.4M NaCl salinity or high irradiance of 1000 micromol m(-2)s(-1), as well as to both stress factors combined (LLSA=low light plus salt; HLCO=high light of 1000 micromol m(-2)s(-1), no salt; HLSA=high light plus salt). A control of LLCO=low light of 200 micromol m(-2)s(-1), no salt was used. Parameters of chlorophyll a fluorescence of photosystem II (PSII) were measured with a pulse amplitude modulated fluorometer. HLCO and LLSA conditions induced a weak degree of CAM with day/night changes of malate levels (Deltamalate) of approximately 12mM in the course of the experiment, while HLSA induced stronger CAM of Deltamalate approximately 20 mM. Effective quantum yield of PSII, DeltaF/F'(m), was only slightly affected by LLSA, somewhat reduced during the course of the experiment by HLCO and clearly reduced by HLSA. Potential quantum efficiency of PSII, F(v)/F(m), at predawn times was not affected by any of the conditions, always remaining at 0.8, showing that there was no acute photoinhibition. During the course of the days HL alone (HLCO) also did not elicit photoinhibition; salt alone (LLSA) caused acute photoinhibition which was amplified by the combination of the two stresses (HLSA). Non-photochemical, NPQ, quenching remained low (<0.5) under LLCO, LLSA and HLCO and increased during the course of the experiment under HLSA to 1-2. Maximum apparent photosynthetic electron transport rates, ETR(max), declined during the daily courses and were reduced by LLSA and to a similar extent by HLSA. It is concluded that M. crystallinum expresses effective stress tolerance mechanisms but photosynthetic capacity is reduced by the synergistic effects of salinity and light irradiance stress combined.
Assuntos
Clorofila/metabolismo , Luz , Mesembryanthemum/metabolismo , Cloreto de Sódio/farmacologia , Clorofila/efeitos da radiação , Fluorescência , Malatos/metabolismo , Mesembryanthemum/efeitos dos fármacos , Mesembryanthemum/efeitos da radiação , Fotossíntese , Complexo de Proteína do Fotossistema II/metabolismoRESUMO
One of the limits of Cd2+-phytoextraction is the high toxicity of this metal to plants. Growth restriction, chlorosis and necrosis are usually accompanied with a large disturbance of the uptake of essential elements. This work aims to study the effects of cadmium (Cd2+) on potassium (K+), calcium (Ca2+) and nitrogen (N) acquisition, and their consequences on growth in two halophytes species: Sesuvium portulacastrum and Mesembryanthemum crystallinum. Seedlings were grown for 30 days in split-root conditions. One half of the root system was immersed in complete nutrient solution (Basal medium (B)) supplemented with 100 microM Cd2+, and the other half was immersed in a Cd2+-free medium, containing all nutrients (B/Cd plants) or deprived of potassium ((B-K)/Cd) or calcium ((B-Ca)/Cd) or nitrogen ((B-N)/Cd). Using this approach, we demonstrated that K+ and Ca2+ uptake was impaired in roots exposed to Cd2+. Concerning N, we noticed no indication of uptake inhibition by Cd2+. However, restriction of K+ uptake by roots was compensated by an increase in the K+-use efficiency, so that growth was not inhibited. Calcium uptake was strongly limited by Cd2. This inhibition was accompanied by a reduction in growth of ((B-Ca)/Cd) plants. Thus, we conclude that Cd2+ limits growth of both halophytes through restriction imposed on Ca2+ uptake. We suggest that the increase of Ca2+ availability in soils could improve the growth of both species in the presence of Cd2+. This would be essential for improving their utility for extraction of this metal by from salty contaminated soils.
Assuntos
Aizoaceae/efeitos dos fármacos , Aizoaceae/metabolismo , Cádmio/toxicidade , Cálcio/metabolismo , Mesembryanthemum/efeitos dos fármacos , Nitrogênio/metabolismo , Potássio/metabolismo , Aizoaceae/crescimento & desenvolvimento , Mesembryanthemum/crescimento & desenvolvimento , Mesembryanthemum/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Plântula/metabolismoRESUMO
The relative influence of plant age and environmental stress signals in triggering a shift from C(3) photosynthesis to Crassulacean acid metabolism (CAM) in the annual halophytic C(3)-CAM species Mesembryanthemum crystallinum was explored by continuously monitoring net CO(2) exchange of whole shoots from the seedling stage until seed set. Plants exposed to high salinity (400 mm NaCl) in hydroponic culture solution or grown in saline-droughted soil acquired between 11% and 24% of their carbon via net dark CO(2) uptake involving CAM. In contrast, plants grown under nonsaline, well-watered conditions were capable of completing their life cycle by operating in the C(3) mode without ever exhibiting net CO(2) uptake at night. These observations are not consistent with the widely expressed view that the induction of CAM by high salinity in M. crystallinum represents an acceleration of preprogrammed developmental processes. Rather, our study demonstrates that the induction of the CAM pathway for carbon acquisition in M. crystallinum is under environmental control.
Assuntos
Dióxido de Carbono/metabolismo , Meio Ambiente , Mesembryanthemum/metabolismo , Fotossíntese/fisiologia , Mesembryanthemum/efeitos dos fármacos , Mesembryanthemum/crescimento & desenvolvimento , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Transpiração Vegetal/fisiologia , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Plântula/metabolismo , Cloreto de Sódio/farmacologia , Água/metabolismoRESUMO
Changes in lipid content and fatty acid composition were determined in leaves of two halophytes: Sesuvium portulacastrum and Mesembryanthemum crystallinum exposed to cadmium (Cd). Experiments were carried out using young small-sized plants grown hydroponically (S. portulacastrum) or aseptically germinated seeds (M. crystallinum). Cd treatment was applied at different concentrations (0, 50, 100 and 200microM) for 30 days. At high cadmium doses (200microM), contents of total lipids (TL) and lipid fractions including galactolipids (GL), phospholipids (PL) and neutral lipids (NL) decreased more in M. crystallinum leaves than in S. portulacastrum leaves. Moreover, there were no significant changes in the total fatty acid composition of S. portulacastrum leaves during metal treatment. In contrast, M. crystallinum leaves showed a decrease in the percentage of the tri-unsaturated fatty acid (C18:3), and a corresponding increase in the percentage of di-unsaturated fatty acid (C18:2). These different responses suggested that S. portulacastrum seems to be more feasible for phytoremediation.
Assuntos
Aizoaceae/química , Aizoaceae/efeitos dos fármacos , Cádmio/farmacologia , Ácidos Graxos/análise , Lipídeos/análise , Mesembryanthemum/química , Mesembryanthemum/efeitos dos fármacosRESUMO
Under salt stress conditions, the level of CpNpG-methylation (N is any nucleoside) of the nuclear genome of the facultative halophyte Mesembryanthemum crystallinum in the CCWGG sequences (W = A or T) increases two-fold and is coupled with hypermethylation of satellite DNA on switching-over of C3-photosynthesis to the crassulacean acid metabolism (CAM) pathway of carbon dioxide assimilation. The methylation pattern of the CCWGG sequences is not changed in both the 5'-promoter region of the gene of phosphoenolpyruvate carboxylase, the key enzyme of C4-photosynthesis and CAM, and in the nuclear ribosomal DNA. Thus, a specific CpNpG-hypermethylation of satellite DNA has been found under conditions of expression of a new metabolic program. The functional role of the CpNpG-hypermethylation of satellite DNA is probably associated with formation of a specialized chromatin structure simultaneously regulating expression of a large number of genes in the cells of M. crystallinum plants on their adaptation to salt stress and switching-over to CAM metabolism.
Assuntos
Adaptação Biológica , Metilação de DNA , DNA de Plantas/química , Regulação da Expressão Gênica de Plantas , Mesembryanthemum/genética , Cloreto de Sódio/farmacologia , Adaptação Biológica/efeitos dos fármacos , Adaptação Biológica/fisiologia , Sequência de Bases , DNA de Plantas/metabolismo , Genoma de Planta , Mesembryanthemum/efeitos dos fármacos , Mesembryanthemum/metabolismo , Dados de Sequência Molecular , Sementes/citologia , Sementes/metabolismo , Cloreto de Sódio/metabolismoRESUMO
In the halophytic species Mesembryanthemum crystallinum, crassulacean acid metabolism (CAM) may be induced by a range of abiotic factors including drought, salinity, high light intensity, low temperature, and anoxia. A key biotic consequence of all these environmental changes is the generation of reactive oxygen species in planta that can elicit potentially damaging oxidative reactions and/or act as signals for engaging mechanisms that alleviate oxidative stress. However, induction of CAM per se also has the potential for increasing the oxidative burden via the enhanced internal O2 concentrations that develop behind closed stomata during daytime decarboxylation. The aim of this paper was to test two hypotheses. The first one, that reactive oxygen species are key signals for up-regulating the major genes and proteins required for the operation of CAM as part of an integrated strategy for alleviating oxidative burden, was tested using gaseous ozone to increase the oxidative burden at a cellular level. The second hypothesis, that CAM potentially increases oxidative load, was tested using a CAM-deficient mutant of M. crystallinum. The data indicate that ozone, like salinity, elicits an increase in the transcript and protein abundance of myo-inositol o-methyl transferase (a key enzyme of cyclitol synthesis), together with phosphoenolpyruvate carboxylase and other 'CAM-related' enzymes. However, ozone, unlike salinity, does not induce functional CAM, implying that the various metabolic components required for CAM respond to different signals. Comparing the activities of different subcellular isoforms of superoxide dismutase in wild-type and CAM-deficient mutants of M. crystallinum suggests that the induction of CAM potentially curtails the oxidative load in planta.
Assuntos
Regulação da Expressão Gênica de Plantas , Mesembryanthemum/metabolismo , Estresse Oxidativo , Proteínas de Plantas/metabolismo , Mesembryanthemum/efeitos dos fármacos , Mesembryanthemum/genética , Metiltransferases/metabolismo , Ozônio/farmacologia , Fosfoenolpiruvato Carboxilase/metabolismo , Fotossíntese , Proteínas de Plantas/genética , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Cloreto de Sódio/farmacologia , Superóxido Dismutase/metabolismoRESUMO
Growth, cadmium accumulation and potassium and calcium status were studied in two halophytes from Aizoaceae family: Sesuvium portulacastrum and Mesembryanthemum crystallinum. After multiplication, the seedlings were cultivated on nutrient solution supplemented with NaCl (100mM) and CdCl2 (0, 50, 100, 200 and 300 microM). After 1 month of treatment, plants were harvested and the dry weight, as well as the Cd, K and Ca concentrations in tissues were determined. Results showed that S. portulacastrum, a perennial halophyte with slow growth, is significantly more tolerant to Cd than M. crystallinum, an annual plant. Cd severely inhibited Mesembryanthemum growth even at the lowest Cd concentration in culture medium (50 microM), and did not modify significantly that of Sesuvium. For both halophytes, Cd accumulation was significantly higher in the roots than in the shoots. However, Cd concentration reached 350-700 microg g(-1) DM in the shoots, values characteristic of Cd hyperaccumulator plants. The addition of Cd in the culture medium led to a disturbance of Ca and especially K nutrition, suggesting the possibility to improve plant growth and Cd phytoextraction of both halophytes by increasing nutrient availability in the culture medium.
Assuntos
Aizoaceae/efeitos dos fármacos , Cloreto de Cádmio/farmacologia , Mesembryanthemum/efeitos dos fármacos , Aizoaceae/crescimento & desenvolvimento , Aizoaceae/metabolismo , Meios de Cultura , Mesembryanthemum/crescimento & desenvolvimento , Mesembryanthemum/metabolismoRESUMO
The carbon isotope composition of the halophyte Mesembryanthemum crystallinum L. (Aizoaceae) changes when plants are exposed to environmental stress and when they shift from C(3) to crassulacean acid metabolism (CAM). We examined the coupling between carbon isotope composition and photosynthetic pathway by subjecting plants of different ages to salinity and humidity treatments. Whole shoot delta(13)C values became less negative in plants that were exposed to 400 mM NaCl in the hydroponic solution. The isotopic change had two components: a direct NaCl effect that was greatest in plants still operating in the C(3) mode and decreased proportionally with increasing levels of dark fixation, and a second component related to the degree of CAM expression. Ignoring the presumably diffusion-related NaCl effect on carbon isotope ratios results in an overestimation of nocturnal CO(2) gain in comparison to an isotope versus nocturnal CO(2) gain calibration established previously for C(3) and CAM species grown under well-watered conditions. It is widely taken for granted that the shift to CAM in M. crystallinum is partially under developmental control and that CAM is inevitably expressed in mature plants. Plants, cultivated under non-saline conditions and high relative humidity (RH) for up to 63 days, maintained diel CO(2) gas-exchange patterns and delta(13)C values typical of C(3) plants. However, a weak CAM gas-exchange pattern and an increase in delta(13)C value were observed in non-salt-treated plants grown at reduced RH. These observations are consistent with environmental control rather than developmental control of the induction of CAM in mature M. crystallinum under non-saline conditions.