Salicylic Acid and Calcium Chloride Seed Priming: A Prominent Frontier in Inducing Mineral Nutrition Balance and Antioxidant System Capacity to Enhance the Tolerance of Barley Plants to Salinity.

The current investigation aims to underline the impact of salicylic acid or calcium chloride seed pre-treatments on mineral status and oxidative stress markers, namely levels of electrolyte leakage (EL) and lipid peroxidation levels, measured as thiobarbituric reactive substances (TBARS), and the activity of some antioxidant enzymes in roots and leaves of plants in two barley species grown under various salt treatments. Overall, our results revealed that salinity inhibits essential nutrient absorption such as iron, calcium, magnesium and potassium and stimulates the absorption of sodium. Also, this environmental constraint induced oxidative stress in plants in comparison with the control conditions. This state of oxidative stress is reflected by an increase in TBARS content as well as the stimulation of EL values. In addition, salinity induced disturbances in the activity of antioxidant enzymes, which were mainly dependent on the applied salt concentration and the species. In addition, Hordeum marinum maintained high antioxidant enzyme activity and low levels of oxidative stress parameters, which reinforces its salt-tolerant character. Importantly, salicylic acid or calcium chloride seed priming alleviated the mineral imbalance and the oxidative damage induced by salinity. Moreover, seed priming improves iron, calcium magnesium and potassium content and limitsthe accumulation of sodium. Also, both treatments not only decrease TBARS levels and limit EL, but they also stimulate the antioxidant enzyme activities in the leaves and roots of the stressed plants as compared with stressed plants grown from non-primed seeds. Interestingly, the beneficial effects of the mentioned treatments were more notable on Hordeum vulgare species.

Silicon (Si) mitigates the negative effects of iron deficiency in common bean (Phaseolus vulgaris L.) by improving photosystem activities and nutritional status.

Silicon (Si) is the second most abundant element in the Earth's crust after oxygen. Its beneficial impact on crop development and yield, particularly under stressful conditions such as iron (Fe) deficiency, has been well documented. Fe deficiency is a critical constraint that limits crop production globally. The objective of this study was to investigate the effects of silicon (Na2SiO3) on common bean (Phaseolus vulgaris L. 'Coco Rose' variety) under iron-deficient conditions. The common bean plants were subjected to six treatments, which included three sufficient iron treatments (50 µM Fe) each paired with three varying silicon concentrations (0, 0.25, and 0.5 mM Si), and three iron-deficient treatments (0.1 µM Fe) each associated with the same silicon concentrations (0, 0.25, and 0.5 mM Si). The results indicate that iron deficiency had a negative impact on almost all the measured parameters. However, under silicon treatments, especially with 0.5 mM Si, the depressive effects of iron deficiency were significantly mitigated. The addition of 0.5 mM Si alleviated leaf chlorosis and improved biomass production, nutritional status, photosynthetic pigment content, photosynthetic gas exchange, and photosystem (PSI and PSII) activities. Interestingly, a greater beneficial effect of silicon was observed on PSII compared to PSI. This was accompanied by a significant augmentation in leaf iron concentration by 42%. Therefore, by enhancing the photosystem activities and nutritional status, among other mechanisms, silicon is capable of mitigating the adverse effects of iron-deficient conditions, making it a successful and effective solution to cope with this nutritional stress.

Phytochemical investigation, antioxidant, anti-inflammatory and cytotoxic activities of Tunisian medicinal Tamarix africana Poir.

The current study aimed to evaluate Tunisian Tamarix africana Poir biological activities. In this study, novel biological activities of the shoot extracts related to their phenolics investigated. Secondary metabolite contents, antioxidant, anti-inflammatory and cytotoxic activities of four extracts (hexane, dichloromethane, methanol and water) were investigated. Antioxidant activities were assessed via in vitro and ex vivo assays. Besides, anticancer activity was investigated against human lung carcinoma (A-549) and colon adenocarcinoma (DLD-1) cells. The anti-inflammatory ability was evaluated via inhibition of LPS-induced NO production in RAW 264.7 macrophage cell lines. Methanol and water extracts displayed the highest antioxidant (IC50 = 3.3 and 4.3 µg/mL respectively), which are correlated activities correlated with phenolic contents. Hexane extract exhibited an important anti-inflammatory effect inhibiting NO ability by 100% at 80 µg/mL. Besides, T. africana extracts were found to be active against A-549 lung carcinoma cells with IC50 values ranging from 20 to 34 µg/mL. These results suggested that T. africana is considered as a potential source of readily accessible natural molecules with a promising effect on human health and diseases.

Priming seeds with salicylic acid modulates membrane integrity, antioxidant defense, and gene expression in Medicago sativa grown under iron deficiency and salinity.

Exposure of plants to adverse environmental conditions reduces their growth and productivity. Currently, seed priming with phytohormones is considered one of the most reliable and cost-effective approaches that can help alleviate the toxic effects of environmental stress. In this context, the present study aims to investigate the effect of priming alfalfa seeds with salicylic acid (SA) on oxidative stress markers, including malonyldialdehyde, protein content, activities of antioxidant enzymes, and expression of genes encoding these enzymes in leaves and roots of alfalfa (Gabes ecotype) grown under saline stress, iron deficiency, or both. Our results showed that the application of salt stress and iron deficiency separately or simultaneously induces changes in the activities of antioxidant enzymes, but these are organ- and stress-dependent. The Gabes ecotype was able to increase the activities of these enzymes under salt stress to alleviate oxidative damage. Indeed, priming seeds with 100 µM SA significantly increases the enzymatic activities of APX, GPX, CAT, and SOD. Therefore, this concentration can be considered optimal for the induction of iron deficiency tolerance. Our results showed not only that Gabes ecotype was able to tolerate salt stress by maintaining high expression of the Fe-SOD isoform, but also that the pretreatment of seeds with 100 µM SA improved the tolerance of this ecotype to iron deficiency by stimulating Fe-SOD expression and inhibiting CAT and APXc.

The halotolerant rizhobacterium Glutamicibacter sp. alleviates salt impact on Phragmites australis by producing exopolysaccharides and limiting plant sodium uptake.

Salinity is a widespread abiotic stress, which has strong adverse effects on plant growth and crop productivity. Exopolysaccharides (EPS) play a crucial role in plant growth-promoting rhizobacteria (PGPR)-mediated improvement of plant stress tolerance. This study aimed to assess whether Glutamicibacter sp. strain producing large amounts of EPS may promote tolerance of common reed, Phragmites australis (Cav.) Trin. ex Steud., towards salt stress. This halotolerant rizhobacterium showed tolerance to salinity (up to 1 M NaCl) when cultivated on Luria-Bertani (LB) medium. Exposure to high salinity (300 mM NaCl) significantly impacted the plant growth parameters, but this adverse effect was mitigated following inoculation with Glutamicibacter sp., which triggered higher number of leaves and tillers, shoot fresh weight/dry weight, and root fresh weight as compared to non-inoculated plants. Salt stress increased the accumulation of malondialdehyde (MDA), polyphenols, total soluble sugars (TSSs), and free proline in shoots. In comparison, the inoculation with Glutamicibacter sp. further increased shoot polyphenol content, while decreasing MDA and free proline contents. Besides, this bacterial strain increased tissue Ca+ and K+ content concomitant to lower shoot Na+ and root Cl- accumulation, thus further highlighting the beneficial effect of Glutamicibacter sp. strain on the plant behavior under salinity. As a whole, our study provides strong arguments for a potential utilization of EPS-producing bacteria as a useful microbial inoculant to alleviate the deleterious effects of salinity on plants.

Plant Growth-Promoting Rhizobacteria Alleviate High Salinity Impact on the Halophyte Suaeda fruticosa by Modulating Antioxidant Defense and Soil Biological Activity.

Plant growth-promoting rhizobacteria (PGPR) are considered as bio-ameliorators that confer better salt resistance to host plants while improving soil biological activity. Despite their importance, data about the likely synergisms between PGPR and halophytes in their native environments are scarce. The objective of this study was to assess the effect of PGPR (Glutamicibacter sp. and Pseudomonas sp.) inoculation on biomass, nutrient uptake, and antioxidant enzymes of Suaeda fruticosa, an obligate halophyte native in salt marshes and arid areas in Tunisia. Besides, the activity of rhizospheric soil enzyme activities upon plant inoculation was determined. Plants were grown in pots filled with soil and irrigated with 600 mM NaCl for 1 month. Inoculation (either with Pseudomonas sp. or Glutamicibacter sp.) resulted in significantly higher shoot dry weight and less accumulation of Na+ and Cl- in shoots of salt-treated plants. Glutamicibacter sp. inoculation significantly reduced malondialdehyde (MDA) concentration, while increasing the activity of antioxidant enzymes (superoxide dismutase; catalase; ascorbate peroxidase; and glutathione reductase) by up to 100%. This provides strong arguments in favor of a boosting effect of this strain on S. fruticosa challenged with high salinity. Pseudomonas sp. inoculation increased shoot K+ and Ca2+ content and lowered shoot MDA concentration. Regarding the soil biological activity, Pseudomonas sp. significantly enhanced the activities of three rhizospheric soil enzymes (urease, ß-glucosidase, and dehydrogenase) as compared to their respective non-inoculated saline treatment. Hence, Pseudomonas sp. could have a great potential to be used as bio-inoculants in order to improve plant growth and soil nutrient uptake under salt stress. Indole-3-acetic acid concentration in the soil increased in both bacterial treatments under saline conditions, especially with Glutamicibacter sp. (up to +214%). As a whole, Glutamicibacter sp. and Pseudomonas sp. strains are promising candidates as part of biological solutions aiming at the phytoremediation and reclamation of saline-degraded areas.

Nutraceutical Potential of Leaf Hydro-Ethanolic Extract of the Edible Halophyte Crithmum maritimum L.

Aromatic halophytes represent an exceptional source of natural bioactive compounds for the food industry. Crithmum maritimum L., also known as sea fennel, is a halophyte plant colonizing cliffs and coastal dunes along Mediterranean and Atlantic coasts. It is well known to produce essential oils and polyphenols endowed with antioxidant and biological effects. The present work reports the phytochemical profile, as well as antioxidant, antimicrobial and antimutagenic properties of C. maritimum leaf hydro-alcoholic extract. From LC-ESI-MS analysis, eighteen phenolic compounds were depicted in sea fennel extract and the amount of total phenolic content exceeds 3% DW. Accordingly, C. maritimum extract showed strong antioxidant activities, as evidenced by in vitro (DPPH, ORAC, FRAP) and ex vivo (CAA-RBC and hemolysis) assays. An important antimicrobial activity against pathogenic strains was found as well as a strong capacity to inhibit Staphylococcus aureus (ATCC 35556) biofilm formation. Sea fennel extracts showed a significant decrease of mutagenesis induced by hydrogen peroxide (H2O2) and menadione (ME) in Saccharomyces cerevisiae D7 strain. In conclusion, our results show that C. maritimum is an exceptional source of bioactive components and exert beneficial effects against oxidative or mutagenic mechanisms, and pathogenic bacteria, making it a potential functional food.

Changes in organic compounds secreted by roots in two Poaceae species (Hordeum vulgare and Polypogon monspenliensis) subjected to iron deficiency.

Despite their economic and ecological interests, Poaceae are affected by the low availability of iron in calcareous soils. Several studies focused on the capacity of this family to secrete phytosiderophores and organic acids as a mechanism of tolerance to iron deficiency. This work aimed at studying the physiological responses of two Poaceae species; Hordeum vulgare (cultivated barley) and Polypogon monspenliensis (spontaneous species) to iron deficiency, and evaluate especially the release of phytosiderophores and organic acids. For this purpose, seedlings of these two species were cultivated in complete nutrient solution with or without iron. The biomass production, iron status, phytosiderophores and organic acids release by roots were studied. The results demonstrated that Polypogon monspenliensis was relatively more tolerant to iron deficiency than Hordeum vulgare. Polypogon monspenliensis had the ability to secrete a higher amount of phytosiderophores and organic acids, especially citric, acetic, oxalic and malic acids, compared to Hordeum vulgare. We propose this spontaneous species as a forage plant in calcareous soils and in intercropping systems with fruit trees to prevent iron chlorosis.

Early effects of salt stress on the physiological and oxidative status of the halophyte Lobularia maritima.

Soil salinity is an abiotic stress that reduces agricultural productivity. For decades, halophytes have been studied to elucidate the physiological and biochemical processes involved in alleviating cellular ionic imbalance and conferring salt tolerance. Recently, several interesting genes with proven influence on salt tolerance were isolated from the Mediterranean halophyte Lobularia maritima (L.) Desv. A better understanding of salt response in this species is needed to exploit its potential as a source of stress-related genes. We report the characterisation of L. maritima's response to increasing NaCl concentrations (100-400 mM) at the physiological, biochemical and molecular levels. L. maritima growth was unaffected by salinity up to 100 mM NaCl and it was able to survive at 400 mM NaCl without exhibiting visual symptoms of damage. Lobularia maritima showed a Na+ and K+ accumulation pattern typical of a salt-includer halophyte, with higher contents of Na+ in the leaves and K+ in the roots of salt-treated plants. The expression profiles of NHX1, SOS1, HKT1, KT1 and VHA-E1 in salt-treated plants matched this Na+ and K+ accumulation pattern, suggesting an important role for these transporters in the regulation of ion homeostasis in leaves and roots of L. maritima. A concomitant stimulation in phenolic biosynthesis and antioxidant enzyme activity was observed under moderate salinity, suggesting a potential link between the production of polyphenolic antioxidants and protection against salt stress in L. maritima. Our findings indicate that the halophyte L. maritima can rapidly develop physiological and antioxidant mechanisms to adapt to salt and manage oxidative stress.

Nickel tolerance and toxicity mechanisms in the halophyte Sesuvium portulacastrum L. as revealed by Ni localization and ligand environment studies.

Halophytes are able to tolerate relatively high concentrations of hazardous metals in a growing substrate, what makes them suitable candidates for phytoremediation of metal-contaminated soils. In this work, we aimed to study the physiological responses of the halophyte Sesuvium portulacastrum L. to Ni, with main focus on Ni localization, compartmentation and ligand environment, to decipher Ni tolerance and toxicity mechanisms. Seedlings were grown in hydroponic nutrient solution containing 0, 25, 50 and 100 µM Ni as NiCl2 for 3 weeks. Ni localization in leaves was assessed by micro-proton-induced X-ray emission (micro-PIXE). Ni ligand environment was studied by Ni K-edge X-ray absorption near edge structure (XANES). In addition, Ni-soluble, weakly bound/exchangeable and insoluble leaf tissue fractions were determined by sequential extraction. Results show that S. portulacastrum is able to tolerate up to ~ 500 µg g-1 dry weight (DW) of Ni in the shoots without significant growth reduction. At higher Ni concentrations (> 50 µM Ni in nutrient solution), chloroses were observed due to the accumulation of Ni in photosynthetically active chlorenchyma as revealed by micro-PIXE. Water storage tissue represented the main pool for Ni storage. Incorporation of Ni into Ca-oxalate crystals was also observed in some specimens, conferring tolerance to high leaf Ni concentrations. The majority of Ni (> 70%) was found in soluble tissue fraction. Ni K XANES revealed Ni bound mainly to O- (55%) and N-ligands (45%). Ni toxicity at higher Ni levels was associated with Ni binding to amino groups of proteins in cytosol of chlorenchyma and increased level of lipid peroxidation. Proline levels also increased at high Ni exposures and were associated with Ni-induced oxidative stress and alteration of water regime.

Phytochemical and Biological Activities in Limonium Species Collected in Different Biotopes of Tunisia.

A particular interest is nowadays given to natural antioxidants occurring in foods which can reduce the risk of several diseases through their protective effect. The genus Limonium is widely distributed in different salt regions of Tunisia and known in traditional medicine for the presence of highly effective viral and bacterial replication inhibitors. Limonium leaves have possible beneficial effects on human health for their antioxidant activities and free radical scavenging abilities. To exploit the potential of plants from extreme environments as new sources of natural antioxidants, we studied the extracts from leaves of eight Limonium species growing in extreme environments in Tunisia. Antioxidant molecules (polyphenols, flavonoids, flavonols, ascorbate, tocopherols), in vitro (DPPH, ORAC) and ex vivo antioxidant potential on human erythrocytes, antioxidant enzymes activities (superoxide dismutase, peroxidases, glutathione reductase) were evaluated to identify the species with the best antioxidant capacity. The results showed variability among the species considered in function of the environmental conditions of their natural biotopes, as for the antioxidants measured. In particular, L. vulgare from Oued Rane biotope, characterized by dryness and high temperatures, was the species with the highest enzymatic activity and antioxidant capacity, making it interesting as possible edible halophyte plant or as food complement.

Genetic analysis of cadmium accumulation in lettuce (Lactuca sativa).

This work characterized mechanisms controlling cadmium (Cd) tolerance and accumulation in lettuce at both the physiological and genetic levels. These traits were evaluated in 18 Lactuca accessions representing a large genetic diversity. Cd tolerance and accumulation in roots and shoots as well as Cd translocation from roots to the shoot varied independently, and with a significant range of variation. Analyses of F1 progenies of crosses between cultivars with contrasted phenotypes showed that high tolerance to Cd, low Cd accumulation and low Cd root-shoot translocation were recessive traits. Results of analyses of F2 progenies of different crosses suggest that root Cd concentration and root-shoot Cd translocation were under a complex genetic determinism involving at least two loci. This work thus revealed that limiting both Cd accumulation and Cd root-shoot translocation in lettuce is possible and depends on recessive loci. Differences in the ability to accumulate Cd in roots in the long term could not be linked to differences in short-term 109Cd uptake into, or efflux from, roots. In contrast, the cultivar with the highest root-shoot Cd translocation was the same in the long term and in the short term, which suggests that this trait relies on processes that are implemented quickly (i.e. in less than three days) after the start of Cd exposure.

Physiological response and mineral elements accumulation pattern in Sesuvium portulacastrum L. subjected in vitro to nickel.

Sesuvium portulacastrum, a halophyte with high tolerance to heavy metals like Cd, Pb and Ni is considered for phytoremediation of metal contaminated saline soils. The tolerance to a selected metal ion could, by hypothesis, be stimulated through in vitro adaptation and regeneration of the plant. Seedlings obtained by in vitro micro-propagation, were exposed to 0, 25 and 50 µM Ni, as NiCl2, in agar-based medium for 30 days. Growth parameters, plant water content, the concentration of photosynthetic pigments, proline and malondialdehyde (MDA) concentrations were determined. Nickel and nutrients distribution in leaves was studied by micro-Proton-Induced-X-ray-Emission (µ-PIXE). The results showed that Ni was mainly accumulated in vascular bundles, next in water storage tissues and chlorenchyma. Ni concentrations in chlorenchyma increased with increasing Ni in culturing medium, in direct relation to decrease of photosynthetic pigments and increase of oxidative stress. As compared to control plants, Ni induced substantial increase in MDA and proline accumulation. Plants exposed to 50 µM Ni accumulated up to 650 µg g-1 of Ni in the shoots, exhibiting chlorosis and necrosis and a drastically reduced plant growth. Perturbations in uptake and distribution of nutrients were observed, inducing mineral deficiency, probably through membrane leakage. The mineral nutrient disturbances induced by Ni could be highly implicated in the restriction of S. portulacastrum development under the acute 50 µM Ni level.

Natural Variation in Physiological Responses of Tunisian Hedysarum carnosum Under Iron Deficiency.

Iron (Fe) is an essential element for plant growth and development. The cultivation of leguminous plants has generated strong interest because of their growth even on poor soils. Calcareous and saline soils with poor mineral availability are wide-spread in Tunisia. In an attempt to select better forage crops adapted to Tunisian soils, we characterized Fe deficiency responses of three different isolates of Hedysarum carnosum, an endemic Tunisian extremophile species growing in native stands in salt and calcareous soil conditions. H. carnosum is a non-model crop. The three isolates, named according to their habitats Karkar, Thelja, and Douiret, differed in the expression of Fe deficiency symptoms like morphology, leaf chlorosis with compromised leaf chlorophyll content and photosynthetic capacity and leaf metal contents. Across these parameters Thelja was found to be tolerant, while Karkar and Douiret were susceptible to Fe deficiency stress. The three physiological and molecular indicators of the iron deficiency response in roots, Fe reductase activity, growth medium acidification and induction of the IRON-REGULATED TRANSPORTER1 homolog, indicated that all lines responded to -Fe, however, varied in the strength of the different responses. We conclude that the individual lines have distinct adaptation capacities to react to iron deficiency, presumably involving mechanisms of whole-plant iron homeostasis and internal metal distribution. The Fe deficiency tolerance of Thelja might be linked with adaptation to its natural habitat on calcareous soil.

Potential of the physiological response of pea plants (Pisum sativum L. ) to iron deficiency (direct or lime- induced) / Potencial da resposta fisiológica das plantas de ervilha (Pisum sativum L. ) à deficiência de ferro (direta ou induzida pelo calcário)

Pea (Pisum sativum L.) is an important food crop in Tunisia, where calcareous soils represents the major limiting factor for agriculture production. In the present study a greenhouse experiment was conducted to assess the effects of direct and bicarbonate- induced iron deficiency on plant growth, chlorophyll fluorescence, photosynthesis, spad index and iron nutrition in two Tunisian pea genotypes (Pisum sativum L.). Plants were grown hydroponically and iron deficiency was induced for 3 weeks. Iron deficiency decreased all the above physiological parameters. The direct Fe deficiency is more drastic than bicarbonate- induced Fe deficiency. A close relationship between plant growth, photosynthesis and SPAD index was observed. Fe use efficiency for plant growth and Fe use efficiency for photosynthesis discriminates clearly the studied genotypes and seems to be the main reason of the tolerance of Kelvedon, as compared to Lincoln.

A ervilha (Pisum sativum L.) é uma cultura alimentar importante na Tunísia, onde os solos calcários representam o principal fator limitante para a produção agrícola. No presente estudo, foi conduzido um experimento em estufa para avaliar os efeitos da deficiência de ferro direta e induzida por bicarbonato sobre o crescimento de plantas, a fluorescência da clorofila, a fotossíntese, o índice SPAD e a nutrição de ferro em dois genótipos de ervilha da Tunísia (Pisum sativum L.). As plantas foram cultivadas hidroponicamente e a deficiência de ferro foi induzida durante 3 semanas. A deficiência de ferro diminuiu todos os parâmetros fisiológicos acima. A deficiência de Fe direta é mais drástica do que a deficiência de Fe induzida por bicarbonato. Observou-se uma estreita relação entre o crescimento das plantas, a fotossíntese e o índice SPAD. A eficiência de uso de Fe para o crescimento de plantas e a eficiência de uso de Fe para a fotossíntese discriminam claramente os genótipos estudados e parecem ser a razão principal da tolerância de Kelvedon, em comparação com a de Lincoln.

Cadmium hampers salt tolerance of Sesuvium portulacastrum.

It is well known that salinity reduces cadmium toxicity in halophytes. However, the possible interference of Cd with the mechanisms of salt tolerance is poorly explored. The aim of this study was to see whether Cd affects salt tolerance mechanisms in the halophyte Sesuvium portulacastrum. S. portulacastrum plants obtained from cuttings were grown in hydroponics for 3 weeks and then exposed to low (0.09 mM) or moderate (200 mM) NaCl concentrations, alone or in combination with 25 µM CdCl2. Microscopy observation revealed two strategies of salt tolerance: euhalophytism and secretion of salt by bladder cells. Cadmium exposure hardly influenced the total leaf Na+ concentrations. However, Cd supply delayed the salt-induced upregulation of AHA1 (plasma membrane H+-ATPase 1) and SOS1 (plasma membrane Na+ transporter "Salt Overly Sensitive 1"), genes that are essential for salt tolerance. Moreover, Cd induced the activation of BADH, coding for betaine aldehyde dehydrogenase, indicating enhanced osmotic stress due to Cd. Sodium-green fluorescence in protoplasts from plants grown with low or high NaCl, alone or in combination with Cd, revealed higher Na+ concentrations in the cytoplasm of Cd-exposed plants. Taken together the results indicate interference of Cd with salt tolerance mechanisms in S. portulacastrum. This may have consequences for the efficient use of halophytes in phytoremediation of Cd-contaminated saline soils.

Nickel tolerance, accumulation and subcellular distribution in the halophytes Sesuvium portulacastrum and Cakile maritima.

It has been shown that halophytes are able to successfully cope with heavy metal toxicity, suggesting their possible use for remediation of metal contaminated soils. In this work, Ni tolerance and accumulation in two halophytes, Sesuvium portulacastrum (L.) L. and Cakile maritima Scop. was investigated. Seedlings of both species were subjected hydroponically during 21 days to 0, 25, 50, and 100 µM of NiCl2. The growth and photosynthesis parameters revealed that S. portulacastrum tolerates Ni better than C. maritima. The photosynthesis activity, chlorophyll content and photosystem II integrity were less impacted in Ni-treated S. portulacastrum as compared to C. maritima, although, Ni accumulated in higher concentrations in the shoots of S. portulacastrum (1050 µg g-1 DW) than in those of C. maritima (550 µg g-1 DW). The subcellular fractionation of Ni in the shoots of both species showed that C. maritima accumulated about 65% of Ni in the soluble fraction, while 28% was associated with the cell walls. In S. portulacastrum 44% of the total cellular Ni was seen in the soluble fraction and 43% was bound to the cell walls. It can be concluded that S. portulacastrum tolerates Ni better than C. maritima, most probably due to a better ability to sequester Ni in the cell walls, restricting its accumulation in the soluble fraction.

New Sulphated Flavonoids from Tamarix africana and Biological Activities of Its Polar Extract.

The phytochemical investigation of Tamarix africana Poir. (Tamaricaceae) shoot polar extract afforded three new sulphated flavonoids, (2S,4R)-5,7,4'-trihydroxyflavan-4-ol 5,7-disulphate (1), (2S)-5,7,4'-trihydroxyflavan 7-O-sulphate (2), and (2S)-naringenin 4'-O-sulphate (3), together with ten known compounds. Their structures were determined by spectroscopic methods including 1D and 2D NMR analysis and HRMS. Biological activities of the polar extract of T. africana shoots related to its phenolic content were also investigated. A high total phenolic content (151.1 mg GAE/g) was found in the methanol shoot extract, which exhibits strong antioxidant activities using the oxygen radical absorbance capacity method and a skin cell-based assay. Moreover, the shoot extract showed significant anti-inflammatory activity, reducing nitric oxide release by 53.5 % at 160 µg/mL in lipopolysaccharide-stimulated RAW 264.7 macrophages. Finally, T. africana shoot extract inhibited the growth of A-549 lung carcinoma cells, with an IC50 value of 34 µg/mL.

Proteomic and functional analysis of proline dehydrogenase 1 link proline catabolism to mitochondrial electron transport in Arabidopsis thaliana.

Proline accumulates in many plant species in response to environmental stresses. Upon relief from stress, proline is rapidly oxidized in mitochondria by proline dehydrogenase (ProDH) and then by pyrroline-5-carboxylate dehydrogenase (P5CDH). Two ProDH genes have been identified in the genome of the model plant Arabidopsis thaliana To gain a better understanding of ProDH1 functions in mitochondria, proteomic analysis was performed. ProDH1 polypeptides were identified in Arabidopsis mitochondria by immunoblotting gels after 2D blue native (BN)-SDS/PAGE, probing them with an anti-ProDH antibody and analysing protein spots by MS. The 2D gels showed that ProDH1 forms part of a low-molecular-mass (70-140 kDa) complex in the mitochondrial membrane. To evaluate the contribution of each isoform to proline oxidation, mitochondria were isolated from wild-type (WT) and prodh1, prodh2, prodh1prodh2 and p5cdh mutants. ProDH activity was high for genotypes in which ProDH, most likely ProDH1, was strongly induced by proline. Respiratory measurements indicate that ProDH1 has a role in oxidizing excess proline and transferring electrons to the respiratory chain.

Antioxidant, anti-inflammatory and anticancer activities of the medicinal halophyte Reaumuria vermiculata.

Reaumuria vermiculata is a xero-halophytic specie widely distributed in the south of Tunisia. In the current study, antioxidant, anti-inflammatory and anticancer activities of Reaumuria vermiculata shoot extracts as well as its phenolic compounds were investigated in different solvent extracts (hexane, dichloromethane, methanol and water). Results showed a strong antioxidant activity, using the ORAC method and a cell based-assay, in methanol extract as well as an important phenolic composition (117.12 mg GAE/g). Hexane and dichloromethane proved an interesting anticancer activity against A-549 lung carcinoma cells, with IC50 values of 17 and 23 µg/ml, respectively. Besides, dichloromethane extract displayed the utmost anti-inflammatory activity, inhibiting NO release over 100 % at 80 µg/ml in LPS-stimulated RAW 264.7. Taken together, these finding suggest that R. vermiculata exhibited an interesting biological activities which may be related to the phenolic composition of this plant. Moreover, the identification of phenolic compounds in R. vermiculata dichloromethane extract using RP-HPLC revealed that myricetin was the major molecule. These results allow us to propose R. vermiculata as a valuable source for bioactive and natural compounds exhibiting interesting biological capacities.