Ameliorative effects of Atriplex crassifolia (C.A.Mey) on pain and inflammation through modulation of inflammatory biomarkers and GC-MS-based metabolite profiling.

Atriplex crassifolia (A. crassifolia) is a locally occurring member of Chenopodiaceae family that has been used in folk medicine for the treatment of joint pain and inflammation. The present study was focused to determine the analgesic and anti-inflammatory potential of the plant. n-hexane (ACNH) and methanol (ACM) extracts of A. crassifolia were evaluated for in vitro anti-inflammatory potential using protein denaturation inhibition assay. In vivo anti-inflammatory potential was determined by oral administration of 250, 500, and 1000 mg/kg/day of extracts against carrageenan and formalin-induced paw edema models. Inflammatory mediators such as TNF-α, IL-10, IL-1ß, NF-kB, IL-4, and IL-6 were estimated in blood samples of animals subjected to formalin model of inflammation. Analgesic activity was determined using acetic acid-induced writhing and tail flick assay model. Phytochemical profiling was done by GC-mass spectrophotometer. The results of in vitro anti-inflammatory activity revealed that both ACNH and ACM displayed eminent inhibition of protein denaturation in concentration-dependent manner. In acute in vivo carrageenan-induced paw edema model, both extracts reduced inflammation at 5th and 6th hour of study (p < 0.05). A. crassifolia extracts exhibited significant inhibition against formalin-induced inflammation with maximum effect at 1000 mg/kg. ACNH and ACM significantly augmented the inflammatory mediators (p < 0.05). Levels of TNF-α, IL-6, IL-1ß, and NF-kB were reduced, while those of IL-4 and IL-10 were upregulated. ACNH displayed maximum analgesic effect at 1000 mg/kg, while ACM showed potent activity at 500 and 1000 mg/kg. The extracts restored the CBC, TLC and CRP toward normal. GC-MS analysis revealed the presence of compounds like n-hexadecanoic acid, Phytol, (9E,11E)-octadecadienoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester, 1-hexacosene, vitamin E, campesterol, stigmasterol, gamma sitosterol in both extracts. These compounds have been reported to suppress inflammation by inhibiting inflammatory cytokines. The current study concludes that A. crassifolia possesses significant anti-nociceptive and anti-inflammatory potential owing to the presence of phytochemicals.

Phytochemical investigation and anti-inflammatory potential of Atriplex leucoclada Boiss.

BACKGROUND: The plant kingdom has long been considered a valuable source for therapeutic agents, however, some plant species still untapped and need to be phytochemically and biologically explored. Although several Atriplex species have been investigated in depth, A. leucoclada, a halophytic plant native to Saudi Arabian desert, remains to be explored for its phytochemical content and biological potentials. Herein, the current study investigated the metabolic content and the anti-inflammatory potential of A. leucoclada. METHODS: Powdered aerial parts of the plant were defatted with n-hexane then the defatted powder was extracted with 80% methanol. n-Hexane extract (ATH) was analyzed using GC-MS, while the defatted extract (ATD) was subjected to different chromatographic methods to isolate the major phytoconstituents. The structures of the purified compounds were elucidated using different spectroscopic methods including advanced NMR techniques. Anti-inflammatory activity of both extracts against COX-1 and COX-2 enzymes were examined in vitro. Molecular docking of the identified compounds into the active sites of COX-1 and COX-2 enzymes was conducted using pdb entries 6Y3C and 5IKV, respectively. RESULTS: Phytochemical investigation of ATD extract led to purification and identification of nine compounds. Interestingly, all the compounds, except for 20-hydroxy ecdysone (1), are reported for the first time from A. leucoclada, also luteolin (6) and pallidol (8) are isolated for the first time from genus Atriplex. Inhibitory activity of ATD and ATH extracts against COX-1 and COX-2 enzymes revealed concentration dependent activity of both fractions with IC50 41.22, 14.40 µg/ml for ATD and 16.74 and 5.96 µg/ml for ATH against COX-1 and COX-2, respectively. Both extracts displayed selectivity indices of 2.86 and 2.80, respectively as compared to 2.56 for Ibuprofen indicating a promising selectivity towards COX-2. Molecular docking study supported in vitro testing results, where purified metabolites showed binding affinity scores ranged from -9 to -6.4 and -8.5 to -6.6 kcal/mol for COX-1 and 2, respectively, in addition the binding energies of GC-MS detected compounds ranged from -8.9 to -5.5 and -8.3 to -5.1 kcal/mol for COX-1 and 2, respectively as compared to Ibuprofen (-6.9 and -7.5 kcal/mol, respectively), indicating high binding affinities of most of the compounds. Analysis of the binding orientations revealed variable binding patterns depending on the nature of the compounds. Our study suggested A. leucoclada as a generous source for anti-inflammatory agents.

Isolation and molecular characterization of an FSK2-type dehydrin from Atriplex halimus.

Dehydrins form the group II LEA protein family and are known to play multiple roles in plant stress tolerance and enzyme protection. They harbor a variable number of conserved lysine rich motifs (K-segments) and may also contain three additional conserved motifs (Y-, F- and S-segments). In this work, we report the isolation and characterization of an FSK2-type dehydrin from the halophytic species Atriplex halimus, which we designate as AhDHN1. In silico analysis of the protein sequence revealed that AhDHN1 contains large number of hydrophilic residues, and is predicted to be intrinsically disordered. In addition, it has an FSK2 architecture with one F-segment, one S-segment, and two K-segments. The expression analysis showed that the AhDHN1 transcript is induced by salt and water stress treatments in the leaves of Atriplex seedlings. Moreover, circular dichroism spectrum performed on recombinant AhDHN1 showed that the dehydrin lacks any secondary structure, confirming its intrinsic disorder nature. However, there is a gain of α-helicity in the presence of membrane-like SDS micelles. In vitro assays revealed that AhDHN1 is able to effectively protect enzymatic activity of the lactate dehydrogenase against cold, heat and dehydration stresses. Our findings strongly suggest that AhDHN1 can be involved in the adaptation mechanisms of halophytes to adverse environments.

Improved salt-tolerance of transgenic soybean by stable over-expression of AhBADH gene from Atriplex hortensis.

KEY MESSAGE: The salt-tolerance of transgenic soybean cleared for environmental release was improved by stable over-expression of AhBADH gene from Atriplex hortensis, which was demonstrated through molecular analysis and field experiments. An effective strategy for increasing the productivity of major crops under salt stress conditions is the development of transgenics that harbor genes responsible for salinity tolerance. Betaine aldehyde dehydrogenase (BADH) is a key enzyme involved in the biosynthesis of the osmoprotectant, glycine betaine (GB), and osmotic balance in plants, and several plants transformed with BADH gene have shown significant improvements in salt tolerance. However, very few field-tested transgenic cultivars have been reported, as most of the transgenic studies are limited to laboratory or green house experiments. In this study, we demonstrated through field experiments that AhBADH from Atriplex hortensis confers salt tolerance when transformed into soybean (Glycine max L.). AhBADH was successfully introduced into soybean by Agrobacterium mediated transformation. A total of 256 transgenic plants were obtained, out of which 47 lines showed significant enhancement of salt tolerance compared to non-transgenic control plants. Molecular analyses of the transgenic line TL2 and TL7 with the highest salt tolerance exhibited stable inheritance and expression of AhBADH in progenies with a single copy insertion. TL1, TL2 and TL7 exhibited stable enhanced salt tolerance and improved agronomic traits when subjected to 300mM NaCl treatment. Currently, the transgenic line TL2 and TL7 with stable enhanced salt tolerance, which have been cleared for environmental release, are under biosafety assessment. TL 2 and TL7 stably expressing AhBADH could then be applied in commercial breeding experiments to genetically improve salt tolerance in soybean.

Phytoremediation Potential and Physiological Mechanisms Underlying Metallic Extraction of Suaeda glauca, Artemisia desertorum, and Atriplex canescens.

Mining activities have led to serious environmental (soil erosion, degradation of vegetation, and groundwater contamination) and human health (musculoskeletal problems, diarrheal conditions, and chronic diseases) issues at desert mining areas in northwest China. Native plant species grown naturally in desert regions show a unique tolerance to arid and semiarid conditions and are potential candidates for soil phytoremediation. Here, an ex situ experiment involving pot planting of seedlings of three native plant species (Suaeda glauca, Artemisia desertorum, and Atriplex canescens) was designed to explore their phytoremediation potential and the underlying physiological mechanism. For Zn and Cu, the three plants were all with a biological accumulation coefficient (BAC) greater than 1. For Cd, Ni, and Pb, Atriplex canescens had the highest bioaccumulation concentrations (521.52, 862.23, and 1734.59 mg/kg), with BAC values (1.06, 1.30, 1.25) greater than 1, which indicates that Atriplex canescens could be a broad-spectrum metal extraction plant. Physiological analysis (antioxidation, extracellular secretions, photosynthesis, and hydraulics) showed that the three desert plants exploited their unique strategy to protect against the stress of complex metals in soils. Moreover, the second growing period was the main heavy metal accumulation and extraction stage concomitant with highest water use efficiency (iWUE). Taken together, the three desert plants exhibited the potent heavy metal extraction ability and physiological and ecological adaptability to a harsh polluted environment in arid desert areas, providing potential resources for the bioremediation of metal-contaminated soils in an arid and semiarid desert environment.

Dry matter intake, digestibility, nitrogen utilization and fermentation characteristics of sheep fed Atriplex hay-based diet supplemented with discarded dates as a replacement for barley grain.

This experiment aimed to investigate the effects of substituting barley grain with discarded dates on intake and digestibility, nitrogen (N) utilization and fermentation characteristics of sheep fed Atriplex hay-based diets. Four Barki sheep (50.9 ± 2.93 kg of body weight (BW)) fitted with rumen cannula were used in a 4 × 4 Latin Square design with 23-day periods. Four isonitrogenous (~140 g crude protein (CP)/kg of dry matter (DM)) experimental diets were formulated with Atriplex hay, and dietary sugar content was increased by replacing barley grain with discarded dates (the proportions of discarded dates in the diet were 0, 93, 187 and 280 g/kg DM). Nutrient intakes were not influenced by diet. Organic matter, CP and neutral detergent fibre digestibility increased (p < 0.05) linearly and DM digestibility tended to increase with increased dates inclusion. Ruminal pH and molar proportion of butyrate increased (p < 0.05) linearly with increasing levels of dates inclusion, whereas ruminal ammonia-N decreased (p < 0.05) linearly. Total volatile fatty acid concentration was unaffected. Microbial protein synthesis (MCP) increased linearly and efficiency of microbial protein synthesis tended to increase as discarded dates increased. Total N intake was not affected by diets whereas, urinary N, urea-N and total N execrations linearly decreased (p < 0.05) with higher amounts of dates in the diets. Blood urea-N tended to decrease with increasing levels of dates. It could be concluded that replacing 100% barley grain with discarded dates as a source of soluble carbohydrates can enhance the utilization of N in Atriplex hay-based diet and consequently improve digestibility and MCP.

Availability of copper in mine tailings with humic substance addition and uptake by Atriplex halimus.

The chemical characteristics of mine tailings, organic amendments (doses), and plants are the critical factors that must be evaluated and monitored to ensure the sustainability of phytostabilization. The aim of this study was to evaluate the mobility of copper (Cu) in mine tailings (MT) of the Zone Central of Chile to which commercial humic substances were added, examining their effect on the uptake of Atriplex halimus. Two commercial humic substances (HS1 and HS2) extracted from leonardite (highly oxidized lignite), of different pH and total organic carbon, were evaluated by adsorption curve for Cu. In columns, soluble Cu, pH, and electrical conductivity in leachates were evaluated for MT, MT + HS1, and MT + HS2, and HS1 and HS2 in doses of 120 mg kg-1. In pot assay, seeds were germinated directly in MT and cultivated for 140 days with the addition of HS2 in 120 and 240 mg kg-1. Mine tailing presents high concentration of Cu (2016 ± 223 mg kg-1, pH 6.3 ± 0.1). The results of sequential extraction indicate that Cu is associated with the sulfide fraction of low risk of mobility. The amount of Cu sorbed by HS1 was higher than that sorbed by HS2, and both humic substances showing better fit to the Freundlich than Langmuir model. Lixiviation of Cu was significantly lower in MT + HS1 (0.166 ± 0.043 mg kg-1) and MT + HS2 (0.157 ± 0.018 mg kg-1) than in MT (0.251 ± 0.052 mg kg-1). Copper concentration in plants reached 185.8 ± 37.8 mg kg-1 in the roots and 32.6 ± 7.4 mg kg-1 in the aerial parts cultivated in MT without effect of the humic substance addition in Cu uptake nor growth. Copper concentrations in the aerial parts were adjusted to sufficient or normal levels in plant. A good management of mine tailings through phytostabilization could consider an adequate mixture of humic substances (to avoid leaching of metals) and an organic amendment that provides essential nutrients and increases biomass generation.

Comparative effects of arsenite (As(III)) and arsenate (As(V)) on whole plants and cell lines of the arsenic-resistant halophyte plant species Atriplex atacamensis.

Whole plants and hypocotyl-derived calli of the halophyte plant species Atriplex atacamensis were exposed to 50 µM arsenate (As(V)) or 50 µM arsenite (As(III)). At the whole plant level, As(III) was more toxic than As(V): it reduced plant growth, stomatal conductance, photosystem II efficiency while As(V) did not. In roots, As accumulated to higher level in response to As(III) than in response to As(V). Within root tissues, both arsenate and arsenite were identified in response to each treatment suggesting that oxidation of As(III) may occur. More than 40% of As was bound to the cell wall in the roots of As(V)-treated plants while this proportion strongly decreased in As(III)-treated ones. In leaves, total As and the proportion of As bound to the cell wall were similar in response to As(V) and As(III). Non-protein thiol increased to higher extent in response to As(V) than in response to As(III) while ethylene synthesis was increased in As(III)-treated plants only. Polyamine profile was modified in a contrasting way in response to As(V) and As(III). At the callus level, As(V) and As(III) 50 µM did not reduce growth despite an important As accumulation within tissues. Calli exposed to 50 µM As did not increase the endogenous non-protein thiol. In contrast to the whole plants, arsenite was not more toxic than arsenate at the cell line level and As(V)-treated calli produced higher amounts of ethylene and malondialdehyde. A very high dose of As(V) (1000 µM) strongly reduced callus growth and lead to non-protein thiols accumulation. It is concluded that As(III) was more toxic than As(V) at the plant level but not at the cellular level and that differential toxicity was not fully explained by speciation of accumulated As. Arsenic resistance in A. atacamensis exhibited a cellular component which however did not reflect the behavior of whole plant when exposed to As(V) or As(III).

Phytoremediation of mine tailings with Atriplex halimus and organic/inorganic amendments: A five-year field case study.

Mine tailings have adverse chemical and physical conditions, including high concentrations of metals and salts, low organic matter content, and unbalanced rates of nutrients which limit the development of vegetation. A large scale field experiment was conducted to reclaim a tailing pond by triggering the growth of native species by spontaneous colonization by tilling (TL) the tailing pond surface and using marble waste (CaCO3; MW), pig slurry (PS) and their combination (MW + PS) as soil amendments. Soil physicochemical properties and water and DTPA extractable metal concentrations of bulk and rhizosphere soils were analyzed after five year from the application of the treatments. In addition, plants of Atriplex halimus from each treatment were collected and metals in roots, leaves and stems analyzed. Before amendments application, the studied pond showed a neutral pH, high salinity and a moderate organic carbon content. After five years, the pH value was significantly increased only in MW plot. The results showed significant increases of DTPA-extractable Zn in MW and MW + PS plots, Pb in all treatments except MW plot, Cd only in PS plot, and Cu only in MW + PS plot. A. halimus was the most dominant species, growing spontaneously in all plots, with lower vegetation cover in CT and MW plots, 6% and 2% respectively. Application of MW increased leaf Pb accumulation by 2.5-fold and Cd by 55%, when compared to the CT. The high initial salinity and probable substitution of metals by Ca2+ on exchangeable surfaces of soil particles may be the reasons for higher uptake of metals in MW plot when compared to the other plots. Although this plant is widely utilized in contaminated sites for phytostabilization purposes, it may absorb and translocate high concentrations of metals to the aboveground tissues in saline contaminated sites.

An Alfin-like gene from Atriplex hortensis enhances salt and drought tolerance and abscisic acid response in transgenic Arabidopsis.

Alfin-like (AL) is a small plant-specific gene family with prominent roles in root growth and abiotic stress response. Here, we aimed to identify novel stress tolerance AL genes from the stress-tolerant species Atriplex hortensis. Totally, we isolated four AhAL genes, all encoding nuclear-localized proteins with cis-element-binding and transrepression activities. Constitutive expression of AhAL1 in Arabidopsis facilitated plants to survive under saline condition, while expressing anyone of the other three AhAL genes led to salt-hypersensitive response, indicating functional divergence of AhAL family. AhAL1 also conferred enhanced drought tolerance, as judged from enhanced survival, improved growth, decreased malonaldehyde (MDA) content and reduced water loss in AhAL1-expressing plants compared to WT. In addition, abscisic acid (ABA)-mediated stomatal closure and inhibition of seed germination and primary root elongation were enhanced in AhAL1-transgenic plants. Further analysis demonstrated that AhAL1 could bind to promoter regions of GRF7, DREB1C and several group-A PP2C genes and repress their expression. Correspondingly, the expression levels of positive stress regulator genes DREB1A, DREB2A and three ABFs were all increased in AhAL1-expressing plants. Based on these results, AhAL1 was identified as a novel candidate gene for improving abiotic stress tolerance of crop plants.

Pressure-volume (P-V) curves in Atriplex nummularia Lindl. for evaluation of osmotic adjustment and water status under saline conditions.

The survival of Atriplex nummularia plants in saline environments is possible mainly due to the presence of salt-accumulating epidermal vesicles. Commonly, destructive methods, such as plant material maceration and subsequent reading in osmometers, are employed in studies on water relations and osmotic adjustment and are inconvenient due to their underestimation of the total water potential inside the cells, which can cause overestimation of an osmotic adjustment that is not present. As a result, methods that preserve leaf structure, such as pressure-volume (P-V) curves, which take into consideration only the salts that compose the symplastic solution, are more adequate. Thus, the main objectives of this study were to evaluate the effect of determination methods of osmotic potential (Ψo) in Atriplex nummularia through destructive and leaf structure-preserving techniques and to determine the water relations of the species under increasing NaCl concentrations. Plants were subjected to daily irrigations, maintaining soil moisture at 80% of field capacity, with solutions of increasing NaCl concentration (0, 0.05, 0.1, 0.2, 0.25 and 0.3 M) for 84 days. Water potential, osmotic potential and osmotic adjustment were determined. In addition, P-V curves were constructed using pressure chambers. Water and osmotic potentials decreased linearly with increasing NaCl concentration in the irrigation solution. The main discrepancies observed were related to the osmotic adjustments determined through maceration and P-V curves. Based on the present research, it was possible to conclude that in studies with species that have salt-accumulating vesicles in the epidermis, such as the plants in the genus Atriplex, constructing P-V curves is more adequate than destructive methods.

Phosphorus deficiency modifies As translocation in the halophyte plant species Atriplex atacamensis.

Most arsenic in surface soil and water exists primarily in its oxidized form, as arsenate (As(V); AsO43-), which is an analog of phosphate (PO43-). Arsenate can be taken up by phosphate transporters. Atriplex atacamensis Phil. is native to northern Chile (Atacama Desert), and this species can cope with high As concentrations and low P availability in its natural environment. To determine the impact of P on As accumulation and tolerance in A. atacamensis, the plants were cultivated in a hydroponic system under four treatments: no As(V) addition with 323µM phosphate (control); 1000µM As(V) addition with 323µM phosphate; no As(V) and no phosphate; 1000µM As(V) addition and no phosphate. Phosphate starvation decreased shoot fresh weight, while As(V) addition reduced stem and root fresh weights. Arsenate addition decreased the P concentrations in both roots and leaves, but to a lesser extent than for P starvation. Phosphorus starvation increased the As concentrations in roots, but decreased it in shoots, which suggests that P deficiency reduced As translocation from roots to shoots. Arsenate addition increased total glutathione, but P deficiency decreased oxidized and reduced glutathione in As(V)-treated plants. Arsenate also induced an increase in S accumulation and nonprotein thiol and ethylene synthesis, and a decrease in K concentrations, effects that were similar for the P-supplied and P-starved plants. In contrast, in As(V)-treated plants, P starvation dramatically decreased total soluble protein content and increased lipid peroxidation, compared to plants supplied with P. Phosphorus nutrition thus appears to be an important component of A. atacamensis response to As toxicity.

The use of spent mushroom compost to enhance the ability of Atriplex halimus to phytoremediate contaminated mine soils.

The mushroom cultivation industry produces a huge amount of spent mushroom compost (SMC), a wide world agricultural organic waste which causes serious environmental problems. However, this cheap organic waste could be useful in the remediation of contaminated soils. The aim of this work was to assess the potential of SMC in combination with the native shrub Atriplex halimus, to phytoremediate two mine soils contaminated with Cd, Pb and Cu. Firstly, to minimize metal availability in the soil, the optimal doses of SMC were determined. Secondly, a phytoremediation assay in greenhouse conditions was carried out to test the effects of A. halimus in combination with SMC at different doses. The results showed the ability of SMC to reduce soil acidity, the mobility of the metals and the enhancement of A. halimus growth. SMC promoted metal immobilization in the root of A. halimus and decreased the translocation from the roots to the shoots. The combination of SMC amendment and A. halimus produced phytostabilization of the metals in the mine soils assayed. In conclusion, SMC represents an adequate organic solid waste which in combination with A. halimus can reduce the adverse impact caused by the high mobility of metals in acid mine soils.

AcEBP1, an ErbB3-Binding Protein (EBP1) from halophyte Atriplex canescens, negatively regulates cell growth and stress responses in Arabidopsis.

An ErbB-3-binding protein gene AcEBP1, also known as proliferation-associated 2G4 gene (PA2G4s) belonging to the M24 superfamily, was obtained from the saltbush Atriplex canescens. Subcellular localization imaging showed the fusion protein AcEBP1-eGFP was located in the nucleus of epidermal cells in Nicotiana benthamiana. The AcEBP1 gene expression levels were up-regulated under salt, osmotic stress, and hormones treatment as revealed by qRT-PCR. Overexpression of AcEBP1 in Arabidopsis demonstrated that AcEBP1 was involved in root cell growth and stress responses (NaCl, osmotic stress, ABA, low temperature, and drought). These phenotypic data were correlated with the expression patterns of stress responsive genes and PR genes. The AcEBP1 transgenic Arabidopsis plants also displayed increased sensitivity under low temperature and evaluated resistance to drought stress. Together, these results demonstrate that AcEBP1 negatively affects cell growth and is a regulator under stress conditions.

Effect of sugarcane vinasse and EDTA on cadmium phytoextraction by two saltbush plants.

Although the use of saltbush plants in metal phytoremediation is well known, there is little information about the impact of sugarcane vinasse (SCV) and EDTA on metal uptake. Heavily cadmium-polluted soil (38 mg kg(-1) Cd) was used in pot and incubation experiments to investigate the Cd phytoextraction potential of wavy saltbush (Atriplex undulata) and quail saltbush (Atriplex lentiformis). EDTA at rates of 3, 6, and 10 mM kg(-1) soil and SCV at rates of 7, 15, and 30 mL kg(-1) soil were added to the polluted soil. The application of EDTA significantly (P = 0.002) reduced the growth of saltbush plants; on the other hand, SCV improved the growth. Both EDTA and SCV increased the availability and root-to-shoot transfer of Cd. The plants of A. lentiformis grown on the soil amended with the highest rate of SCV were able to remove 20.4 % of the total soil Cd during a period of 9 months. Based on the obtained results, it may be concluded that A. lentiformis and sugarcane vinasse could be more effective in the phytoextraction of Cd from the polluted soils.

Molecular and Physiological Mechanisms of Heavy Metal Tolerance in Atriplex halimus.

A study was carried out to identify the mechanisms underlying stress caused by Cd and Pb accumulation in leaves of Atriplex halimus L. collected from habitats representing different kinds of pollution. Mean concentrations of Cd and Pb in aerial parts exceeded the critical levels in polluted plants as compared to reference plants. There were significant reduction in guiacol peroxidase, ascorbate peroxidase and glutathione content in most of polluted plants. The results showed increase in superoxide dismutase enzyme in all polluted plants. The significant increment in catalase enzyme, glutathione S-transeferase and ascorbic acid content were observed in most of polluted plants. Results of the nine differential expressed bands showed down regulation of NADH dehydrogenase and Sedoheptulose-bisphosphatase in polluted plants. In contrast, there were six regulated genes in highly polluted plants, representing transcription factors, membrane transporters and ROS detoxification. The transcription level of phytochelatin synthase showed a significant increase in all polluted plants, while heavy metal ATPase transporter expression significantly increased in some polluted plants. In conclusion, A. halimus may use two different strategies against Cd and Pb stress, in which the molecular and physiological features affords similar levels of Cd and Pb tolerance through binding, sequestration and the reduction of harmful effect of heavy metals.

New septanoside and 20-hydroxyecdysone septanoside derivative from Atriplex portulacoides roots with preliminary biological activities.

The phytochemical investigation of a Tunisian plant Atriplex portulacoides (Chenopodiaceae) led to the isolation of two new compounds designated as portulasoid (2) and septanoecdysone (3) along with the known 20-hydroxyecdysone (20HE) (1). Their chemical structures were elucidated on the basis of extensive spectroscopic methods including ES-HRMS, 1D and 2D-NMR. The isolated compounds were finally tested for their antioxidant activity by using DPPH, ABTS(+), Fe(3+) and catalase assays and also for their antibacterial and anticholinesterase activities.

Differential accumulation of glycinebetaine and choline monooxygenase in bladder hairs and lamina leaves of Atriplex gmelini under high salinity.

Atriplex gmelini is a halophyte and possesses bladder hairs on the leaf surface. It is also known to accumulate the osmoprotectant glycinebetaine (GB). However, it remains unclear whether GB and its biosynthetic enzyme choline monooxygenase (CMO) accumulate in the bladder hairs. Microscopic observation of young leaves showed many bladder hairs on their surfaces, but their total number decreased along with leaf maturity. Sodium Green fluorescent approach revealed Na(+) accumulation in bladder cells of young leaves when A. gmelini was grown at high salinity (250 mM NaCl). Due to fewer bladder hairs in mature leaves, Na(+) accumulation was mostly found in mesophyll cells of mature leaves under high salinity. GB accumulation was found at significant level in both bladder- and laminae-cells without any addition of NaCl and its content increased at high salinity. CMO was not found in bladder hairs or young leaf laminae. Instead, the CMO protein expression was observed in mature leaves and that showed increased accumulation with increasing concentration of NaCl. Furthermore, in situ hybridization experiments revealed the expression of a transporter gene for GB, AgBetT, in the bladder hairs. Based on these results, the synthesis and translocation of GB in A. gmelini were discussed.

Cloning and characterization of a salt responsive gene AcPsbQ1 from Atriplex canescens.

PsbQ is an extrinsic subunit of the photosystem II in eukaryotic photosynthetic organisms. Numerous studies have demonstrated that PsbQ can stabilize the inorganic cofactors and enhance the oxygen release in PSII. The decrease of photosynthesis rate under salinity condition is normally attributed to the high concentration of injurious ions, such as Na(+) and Cl(-), which accumulate in the chloroplast and damage thylakoid membrane under salinity stress. In this study, AcPsbQ1 was isolated from a halophyte Atriplex canescens cDNA library. The AcPsbQ1 contains an open reading frame of 699 bp encoding a 233 amino acid protein. In order to investigate its function, AcPsbQ1 was cloned and transformed into Saccharomyces cerevisiae INVSc1. The heterologous expression of AcPsbQ1 in transgenic yeast significantly helped to increase the adapting and recovery ability of yeast cells under the salt and drought. Quantitative real-time PCR assay was performed to reveal the expression pattern of AcPsbQ1 under different abiotic stresses. On exposure to NaCl stress, the transcript level of AcPsbQ1 was significantly enhanced. AcPsbQ1 expression level was also up-regulated under drought stress. These results indicated that AcPsbQ1 might involve in the response to salt stress in A. canescens.

Impact of compost on metals phytostabilization potential of two halophytes species.

Phytostabilization of heavy metals in contaminated soils should be subject to two conditions, the first is the choice plant must be able to stabilize heavy metals in soil, the second is the plant material which produced from the phytostabilization process must be safe and useful to avoid overload on environmental system. A field experiment was conducted out to evaluate the phytostabilization potential of two halophytes species (Atriplex lentiformis and Atriplex undulata). Compost at rates of 0, 15 and 30 ton ha(-1) was used to examine its role in plant growth and heavy metals uptake. The high rate of compost (30 ton ha(-1)) decreased zinc (Zn) concentrations in the leaves of A. lentiformis and A. undulata by 15.8 and 13.0%, while lead (Pb) in the leaves decreased by 37.6 and 35.2% respectively. Despite the extremely high total heavy metals concentrations in the studied soil, plants of Atriplex were able to grow and maintain shoots metals content below the toxic level and the produced plant materials had a high nutritive value compared to the conventional forage crops. Phosphorus (P) and chloride (Cl) in the roots of Atriplex plants play important function in heavy metals phytostabilization mechanism by the two halophytes plants.