Sodium chloride facilitates the secretohalophyte Atriplex canescens adaptation to drought stress.

Atriplex canescens is a C4 shrub with excellent adaptation to saline and arid environments. Our previous study showed that the secretion of excessive Na+ into leaf salt bladders is a primary strategy in salt tolerance of A. canescens and external 100 mM NaCl can substantially stimulate its growth. To investigate whether NaCl could facilitate Atriplex canescens response to drought stress, five-week-old seedlings were subjected to drought stress (30% of field water capacity) in the presence or absence of additional 100 mM NaCl. The results showed that, under drought stress, the addition of NaCl could substantially improve the growth of A. canescens by increasing leaf relative water content, enhancing photosynthetic activity and inducing a significant declined leaf osmotic potential (Ψs). The addition of NaCl significantly increased Na+ concentration in leaf salt bladders and the Na+ contribution to leaf Ψs, while had no adverse effects on K+ accumulation in leaf laminae. Therefore, the large accumulation of Na+ in salt bladders for enhancing osmotic adjustment (OA) ability is a vital strategy in A. canescens responding to drought stress. In addition, the concentration of free proline, bataine and soluble sugars exhibited a significant increase in the presence of NaCl under drought stress, and the betaine contribution to leaf Ψs was significantly increased by additional NaCl compared with that under drought treatment alone, suggesting that compatible solutes are also involved in OA in addition to functioning as protectants to alleviate water deficit injury.

Terrestrial Toxicity of Synthetic Gas-to-Liquid versus Crude Oil-Derived Drilling Fluids in Soil.

Unlike most other conventional petroleum products that are derived from crude oil, gas-to-liquids (GTLs) are petroleum products that are synthesized from natural gas (methane). This process results in GTL products having no sulfur and low aromatic content, so they should have less impact on human health and the environment compared with crude oil-derived products. The GTLs have been registered for use as nonaqueous base fluids (NABFs) in drilling muds, which aid in the process of drilling wells for oil and gas extraction; it is through these uses and others that they enter terrestrial environments. The primary objective of the present study was to determine whether GTLs were less toxic to terrestrial soil biota than conventional NABFs used for land-based drilling, such as diesel and low-toxicity mineral oil (LTMO). A second objective was to understand the fate and impact of these fluids under more realistic soil and aging conditions of a common west Texas (USA) oil-producing region (i.e., sandy loam soil with low organic matter and a hot arid climate). Acute terrestrial toxicity studies were conducted on the soft-bodied terrestrial invertebrate earthworm (Eisenia fetida) along with 3 plant species-alfalfa (Medicago stavia), thickspike wheatgrass (Elymus lanceolatus), and fourwing saltbrush (Atriplex canescens). We also assessed changes in microbial community structure of the soils following additions of NABF. Overall, the GTL NABFs had lower toxicity compared with conventional NABFs like diesel and LTMO, as measured by invertebrate toxicity, plant seed germination, and impact on the microbial community. Environ Toxicol Chem 2020;39:721-730. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Evaluation of the photosynthetic parameters, emission of volatile organic compounds and ultrastructure of common green leafy vegetables after exposure to non-steroidal anti-inflammatory drugs (NSAIDs).

Understanding the effects of many essential non-steroidal anti-inflammatory drugs (NSAIDs) on plants is still limited, especially at environmentally realistic concentrations. This paper presents the influence of three of the most frequently used NSAIDs (diclofenac, ibuprofen, and naproxen) at environmentally realistic concentrations on the autochthonous green leafy vegetables: orache (Atriplex patula L.), spinach (Spinacia oleracea L.) and lettuce (Lactuca sativa L.). Our research was focused on the determination of the photosynthetic parameters, the emission rate of volatile organic compounds, and the evaluation of the ultrastructure of leaves of studied vegetables after exposure to abiotic stress induced by environmental pollutants, namely NSAIDs. The data obtained indicate a moderate reduction of foliage physiological activity as a response to the stress induced by NSAIDs to the selected green leafy vegetables. The increase of the 3-hexenal and monoterpene emission rates with increasing NSAIDs concentration could be used as a sensitive and a rapid indicator to assess the toxicity of the NSAIDs. Microscopic analysis showed that the green leafy vegetables were affected by the selected NSAIDs. In comparison to the controls, the green leafy vegetables treated with NSAIDs presented irregular growth of glandular trichomes on the surface of the adaxial side of the leaves, less stomata, cells with less cytoplasm, irregular cell walls and randomly distributed chloroplasts. Of the three NSAIDs investigated in this study, ibuprofen presented the highest influence. The results obtained in this study can be used to better estimate the impact of drugs on the environment and to improve awareness on the importance of the responsible use of drugs.

Growth and biochemical changes in quail bush (Atriplex lentiformis (Torr.) S.Wats) under Cd stress.

Halophytes have several advantages to be more effective in metal phytoextraction. Little is known about the Cd-phytoextraction potential of Atriplex lentiformis under different levels of Cd. Seven levels of Cd (0, 40, 80, 120, 160, 200, and 240 mg per kg of soil) were added to A. lentiformis plants grown on pots filled with 5 kg of sandy loam soil. A. lentiformis plants achieve different defense mechanisms to meet the high concentration of Cd in the soil and plant. These mechanisms include reducing the number and area of leaves, minimizing chlorophyll synthesis, and enhancing synthesizing of oxalic acid, phenols, and proline. The critical point of Cd was 9.35 and 183 mg kg-1 for available soil Cd and leaves concentrations, respectively. The maximum level of Cd displayed a 66% decrease in the chlorophyll content of the leaves. On the other hand, the oxalic acid, phenols, and proline in the leaves were increased significantly by 129, 100, and 200% when Cd increased from 0 to 240 mg. The tested plant removed 3.6% of the total soil Cd under the low Cd concentration (40 mg) but under the high level of Cd (240 mg), it only removed a negligible amount of soil Cd (0.74%). The current study confirmed that A. lentiformis plants lost the ability to cleanup Cd from contaminated soil under the high levels of contamination due to the high reduction in the production of dry matter.

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).

Combined effect of Cr-toxicity and temperature rise on physiological and biochemical responses of Atriplex halimus L.

An experiment was conducted to evaluate the combined effect of temperature (26 and 30 °C) and Cr toxicity (0, 100 and 1000 µM Cr) on growth, photosynthesis, water content, Cr and nutrients uptake and translocation. The role of antioxidative enzyme towards stresses tolerance was also investigated. Results showed that the maximum relative growth rate and leaf area per plant of Atriplex halimus L. were recorded at 100 µM Cr and 26 °C. However, presence of Cr reduced net photosynthetic and stomatal conductance rates. Overall, temperature rise enhanced the toxic effect of Cr by reducing growth and photosynthesis and inducing antioxidant enzymes activities. Furthermore, temperature rise increased nutrient uptake, as well as nutrient translocation to aboveground tissues; while it diminished Cr translocation. Finally, roots were the main sink for Cr accumulation in A. halimus. At 1000 µM Cr, root Cr concentrations reached 7.2 and 9.1 mg g-1 at 26 and 30 °C, respectively; while shoot Cr concentrations were 0.45 and 0.44 mg g-1 (26 and 30 °C, respectively). The high Cr-accumulation in roots suggests that A. halimus presents a great potential for phytoremediation, especially phytostabilisation of Cr contaminated soils.

Epidermal bladder cells confer salinity stress tolerance in the halophyte quinoa and Atriplex species.

Epidermal bladder cells (EBCs) have been postulated to assist halophytes in coping with saline environments. However, little direct supporting evidence is available. Here, Chenopodium quinoa plants were grown under saline conditions for 5 weeks. One day prior to salinity treatment, EBCs from all leaves and petioles were gently removed by using a soft cosmetic brush and physiological, ionic and metabolic changes in brushed and non-brushed leaves were compared. Gentle removal of EBC neither initiated wound metabolism nor affected the physiology and biochemistry of control-grown plants but did have a pronounced effect on salt-grown plants, resulting in a salt-sensitive phenotype. Of 91 detected metabolites, more than half were significantly affected by salinity. Removal of EBC dramatically modified these metabolic changes, with the biggest differences reported for gamma-aminobutyric acid (GABA), proline, sucrose and inositol, affecting ion transport across cellular membranes (as shown in electrophysiological experiments). This work provides the first direct evidence for a role of EBC in salt tolerance in halophytes and attributes this to (1) a key role of EBC as a salt dump for external sequestration of sodium; (2) improved K+ retention in leaf mesophyll and (3) EBC as a storage space for several metabolites known to modulate plant ionic relations.

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.

Physiological and biochemical mechanisms preventing Cd-toxicity in the hyperaccumulator Atriplex halimus L.

The xero-halophyte Atriplex halimus L., recently described as Cd-hyperaccumulator, was examined to determine Cd toxicity threshold and the physiological mechanisms involved in Cd tolerance. An experiment was conducted to investigate the effect of cadmium from 0 to 1350 µM on chlorophyll fluorescence parameters, gas exchange, photosynthetic pigment concentrations and antioxidative enzyme activities of A. halimus. Cadmium, calcium, iron, manganese, magnesium, potassium, phosphorous, sodium and zinc concentrations were also analyzed. Plants of A. halimus were not able to survive at 1350 µM Cd and the upper tolerance limit was recorded at 650 µM Cd; although chlorosis was observed from 200 µM Cd. Cadmium accumulation increased with increase in Cd supply, reaching maxima of 0.77 and 4.65 mg g(-1) dry weight in shoots and roots, respectively, at 650 µM Cd. Dry mass, shoot length, specific leaf area, relative growth rate, net photosynthetic rate, stomatal conductance, pigments contents and chlorophyll fluorescence were significantly reduced by increasing Cd concentration. However, the activities of superoxide dismutase (SOD; EC1.15.1.1), catalase (CAT; EC1.11.1.6) and guaiacol peroxidase (GPx; EC1.11.1.7) were significantly induced by Cd. Exposures to Cd caused also a significant decrease in P contents in roots, Mg and Mn contents in shoots and Fe and K contents in roots and shoots and had no effect on Ca, Na and Zn contents. The tolerance of A. halimus to Cd stress might be related with its capacity to avoid the translocation of great amounts of Cd in its aboveground tissues and higher activities of enzymatic antioxidants in the leaf.

Physiological and leaf metabolome changes in the xerohalophyte species Atriplex halimus induced by salinity.

Atriplex halimus is a xerohalophyte plant, which could be used as cash crops. This plant was integrated in Tunisian government programs the aim of which is to rehabilitate saline areas and desert. To investigate its strategies involved in salt tolerance, A. halimus was grown hydroponically under controlled conditions with increasing salinity. Plants were harvested and analyzed after 60 days of treatment. The biomass of A. halimus increased by moderate salinity and decreased significantly at high salinity compared to control plants at 400 mM. Despite of the large amounts of Na(+) observed in the leaves of Atriplex plants, leaf water contents and leaf succulence kept on increasing in treated plants and decreased over 150 mM NaCl. This confirmed the compartmentation and the efficient contribution of Na(+) in the osmotic adjustment. Analysis of the metabolic profiles showed an accumulation of carbohydrates and amino acids. The leaf tissues preferentially stored proline, α alanine and sucrose. Increasing NaCl levels were also accompanied by a significant accumulation of malate in leaves. Involvement of these solutes in osmotic adjustment was considered low. Nevertheless, they seemed to have an important role in controlling photosynthesis which capacity was enhanced by low salinity and decreased with increasing salinity (evaluated by actual photochemical efficiency of photosystem II and chlorophyll contents). The unchanged maximum photochemical efficiency of photosystem II accompanied by the increase of the non-photochemical quenching, the enhancement of the total antioxidant activity and the decrease of the malondialdehyde contents in leaves showed efficient protection of membranes and photosystem II from photo-oxidative damage. This protection seemed to be attributed to proline and sucrose largely accumulated in leaves treated with salt.

Evaluation of Atriplex halimus, Medicago lupulina and Portulaca oleracea for phytoremediation of Ni, Pb, and Zn.

Suitable plant species are able to accumulate heavy metals and to produce biomass useful for non-food purposes. In this study, three endemic Mediterranean plant species, Atriplex halimus, Portulaca oleracea and Medicago lupulina were grown hydroponically to assess their potential use in phytoremediation and biomass production. The experiment was carried out in a growth chamber using half strength Hoagland's solutions separately spiked with 5 concentrations of Pb and Zn (5, 10, 25, 50, and 100 mg L(-1)), and 3 concentrations of Ni (1, 2 and 5 mg L(-1)). Shoot and root biomass were determined and analyzed for their metals contents. A. halimus and M. lupulina gave high shoot biomass with relatively low metal translocation to the above ground parts. Metals uptake was a function of both metals and plant species. It is worth noting that M. lupulina was the only tested plant able to grow in treatment Pb50 and to accumulate significant amount of metal in roots. Plant metal uptake efficiency ranked as follows: A. halimus > M. lupulina > P. oleracea. Due to its high biomass production and the relatively high roots metal contents, A. halimus and M. lupulina could be successfully used in phytoremediation, and in phytostabilization, in particular.

Assessing the effect of copper on growth, copper accumulation and physiological responses of grazing species Atriplex halimus: ecotoxicological implications.

Tolerance of plants to elevated concentrations of heavy metals in growth media and in its tissues leads to high degrees of metal bioaccumulation, which may pose a risk for humans and animals alike. Therefore, bio-accumulating plants need thorough evaluation from an environmental health point of view. A glasshouse experiment concerning the xerohalophyte Atriplex halimus was carried out to determine its tolerance and capacity to accumulate copper. We investigated the effect of Cu from 0 to 30 mmol l(-1) on the growth, photosynthetic apparatus and nutrient uptake of A. halimus by measuring gas exchange, chlorophyll fluorescence and photoinhibition. We also determined total Cu, sodium, potassium, magnesium, phosphorous, and nitrogen content in the plant. Our results indicated that A. halimus presented a high resistance to Cu-induced stress, since the plants were able to survive at concentrations higher than 15 mmol l(-1) Cu. However, this capacity was not reflected in its ability to accumulate and tolerate greater amounts of Cu in its tissues, since clear phytotoxicity symptoms were detected at tissue concentrations greater than 38 mg kg(-1) Cu. Thus, Cu increment caused a reduction in A. halimus growth, which was related to a decrease in net photosynthetic rate. This reduction was associated with the adverse effect of Cu on the photochemical apparatus and the reduction in the absorption of essential nutrients. The high resistance of A. halimus was largely related with the capacity of this species to avoid the absorption of great amounts of Cu. For all the above reasons, A. halimus could have the characteristics of a Cu-exclusion plant.

Plant responses to heterogeneous salinity: growth of the halophyte Atriplex nummularia is determined by the root-weighted mean salinity of the root zone.

Soil salinity is generally spatially heterogeneous, but our understanding of halophyte physiology under such conditions is limited. The growth and physiology of the dicotyledonous halophyte Atriplex nummularia was evaluated in split-root experiments to test whether growth is determined by: (i) the lowest; (ii) the highest; or (iii) the mean salinity of the root zone. In two experiments, plants were grown with uniform salinities or horizontally heterogeneous salinities (10-450 mM NaCl in the low-salt side and 670 mM in the high-salt side, or 10 mM NaCl in the low-salt side and 500-1500 mM in the high-salt side). The combined data showed that growth and gas exchange parameters responded most closely to the root-weighted mean salinity rather than to the lowest, mean, or highest salinity in the root zone. In contrast, midday shoot water potentials were determined by the lowest salinity in the root zone, consistent with most water being taken from the least negative water potential source. With uniform salinity, maximum shoot growth was at 120-230 mM NaCl; ~90% of maximum growth occurred at 10 mM and 450 mM NaCl. Exposure of part of the roots to 1500 mM NaCl resulted in an enhanced (+40%) root growth on the low-salt side, which lowered root-weighted mean salinity and enabled the maintenance of shoot growth. Atriplex nummularia grew even with extreme salinity in part of the roots, as long as the root-weighted mean salinity of the root zone was within the 10-450 mM range.

Arsenic accumulation and distribution in relation to young seedling growth in Atriplex atacamensis Phil.

Even at trace levels, arsenic is of environmental and health concern due to its high toxicity. The xerohalophyte plant species Atriplex atacamensis grows on an arsenic-contaminated mining area in North Chile. Young seedlings that were grown from seeds collected from these plants were grown in a nutrient solution under controlled environmental conditions and were exposed for 14 and 28 days to 0, 100 or 1000 µM arsenate. More than 75% of the plants that were exposed to the highest As dose survived until the end of the treatment. The seedling growth was reduced (100 µM As) or inhibited (1000 µM As) in the stress conditions, but the plants were able to efficiently close their stomata and perform osmotic adjustments to avoid secondary water stress. Arsenic accumulated up to 400 µg g(-1) DW in the shoots and 3500 µg g(-1) DW in the roots. Arsenate drastically impaired the P content and increased glycinebetaine content, although no arsenobetaine was found in the tissues. With the exception of arsenite and arsenate, no As-containing organic compound was detected. Arsenic was not excreted by the trichomes that were present at the leaf surface. Although an increase in the total level of non-protein thiols suggested that arsenite fixation on the sulfhydryl groups could occur in the stressed tissues, the majority of the soluble arsenic remained in its oxidized state As(V). Arsenate induced an increase in the free soluble polyamine concentrations in all of the organs, and it increased the proportion of spermidine and spermine and decreased the proportion of putrescine in the polyamine pool. Therefore, it is likely that these polycationic molecules may assist in arsenate sequestration in the stressed tissues, and A. atacamensis may represent a promising plant species that can be tested in field trials for its phytomanagement of As-contaminated sites in desert areas.

Localization of potential ion transport pathways in vesicular trichome cells of Atriplex halimus L.

The secreting glandular trichomes are recognized as an efficient structure that alleviates salt effects on Atriplex halimus. They are found on buds, young green stems, and leaves. They occupy both the leaf surfaces and give them a whitish color. Their histogenesis and ultrastructure were investigated in the third young leaves. They appear in early stage of plant development and its initiation continuous until just the leaf final development state. Each trichome contains two parts; a stalk which has high electron opacity, embedded in epidermal cells, and bears a second one which is unicellular, called bladder cell and has a low electron density. The bladder cell appears as a huge vacuole and the well-reduced cytoplasm which is pushed close to the wall, contains only a few organelles. Concurrently, the use of silver chloride precipitation technique shows that, in secretion process, salt follows a symplasmatic pathway which is consolidated by the presence of numerous plasmodesmata between the stalk cell(s), and the bladder one and the neighboring mesophyll cells. In addition, according to lanthanum-tracer study, salt can be excreted apoplastically. In fact, the heavy element can be transported via endocytosis vesicles, and by Golgi, endoplasmic reticulum, and lysosome (G.E.R.L.) network toward the storage vacuoles.

Differential responses of saltbush Atriplex halimus L. exposed to salinity and water stress in relation to senescing hormones abscisic acid and ethylene.

Drought and salinity induce water deficit, but may also have distinct effects on plant metabolism. To compare their impact on leaf senescence in relation to ABA and ethylene synthesis, young plants of Atriplex halimus L. were exposed to iso-osmotic concentrations of NaCl (160mM) or PEG (15%) in nutrient solution. Plant growth and development were more affected by PEG than by NaCl. Stressed plants remained able to reduce their osmotic potential, but the nature of accumulated organic osmocompatible solutes varied according to the stressing agent. Glycinebetaine accumulated to a greater extent in salt-treated plants than in water-stressed plants. Sodium chloride induced the accumulation of non-reducing sucrose, while PEG-treated plants mainly accumulated reducing glucose and fructose. Abscisic acid (ABA) accumulated in response to salt, while ethylene was synthesized mainly by PEG-treated plants and was involved in the induction of early senescence processes characterized by synthesis of reactive oxygen species, peroxidation of membrane lipids and a decrease in chlorophyll content. ABA sensitivity of stressed tissues was markedly different in response to salt and in response to non-ionic osmotic stress, and exogenous ABA (50µM) had contrasting effects on most physiological parameters depending on the stressing agent. Exogenous ABA induced a decrease in root and shoot growth and sucrose content, and an increase in reactive oxygen species content in salt-stressed plants. In contrast, exogenous ABA increased growth in PEG-treated plants in relation to an improvement of water use efficiency resulting from a more efficient stomatal control. Exogenous ABA increased ethylene synthesis in salt-treated plants, but had only marginal impact on PEG-treated ones. The xero-halophyte A. halimus thus responds in a contrasting way to salt and water stress, through accumulation of distinct osmocompatible solutes and hormonal compounds such as ethylene and ABA could play distinct roles in stress-induced senescence processes.

Cadmium has contrasting effects on polyethylene glycol-sensitive and resistant cell lines in the Mediterranean halophyte species Atriplex halimus L.

Beside a direct toxicity, cadmium impact on plants involves both a secondary-induced water stress and an oxidative stress. Proliferating cell lines of Atriplex halimus were selected for their sensitivity or resistance to polyethylene glycol (PEG 10,000, 20%) and then exposed to 100 microM CdCl2 in the simultaneous presence or absence of PEG 20% or 150 mM NaCl. The PEG resistant cell line exhibited a higher growth in the presence of Cd than the sensitive line, although Cd acccumulation was higher in the former than in the latter. Exogenous PEG induced an increase in Cd concentration in the sensitive but not in the resistant cell line while NaCl induced a decrease in Cd accumulation in both cell lines. In the presence of Cd alone, the water content (WC) was higher and the osmotic potential was lower in PEG-sensitive than in PEG resistant line. The presence of PEG in the Cd-containing medium increased the WC and decreased the osmotic potential in PEG-resistant line comparatively to Cd stress alone, while an inverse trend was observed for the sensitive line. The PEG-resistant cell line displayed a higher ability to cope with oxidative stress in relation to an increase of endogenous antioxidants (glutathione and ascorbic acid), a high constitutive superoxide dismutase (EC 1.15.1.1) activity and an efficient Cd-induced increase in glutathione reductase (GR) (EC 1.6.4.1) and ascorbate peroxidase (APX) (EC 1.11.1.11). Cadmium tolerance in PEG-resistant line is thus not related to any strategy of Cd exclusion or osmotic adjustment but to tolerance mechanisms allowing the tissue to restrict the deleterious impact of accumulated Cd.

Abscisic acid has contrasting effects on salt excretion and polyamine concentrations of an inland and a coastal population of the Mediterranean xero-halophyte species Atriplex halimus.

BACKGROUND AND AIMS: Different populations of the Mediterranean xerohalophyte species Atriplex halimus exhibit different levels of resistance to salt and osmotic stress depending on the nature of the osmocompatible solute they accumulate. There is, however, no conclusive description of the involvement of abscisic acid (ABA) in the plant response to NaCl or osmotic stress in this species. METHODS: Seedlings issued from an inland water-stress-resistant population (Sbikha) and from a coastal salt-resistant one (Monastir) were exposed in nutrient solutions to NaCl (40 or 160 mm) or to 15 % PEG for 1 d and 10 d in the presence or absence of 50 microm ABA. KEY RESULTS: Plants from Sbikha accumulated higher amounts of ABA in response to osmotic stress than those of Monastir, while an opposite trend was recorded for NaCl exposure. Exogenous ABA improved osmotic stress resistance in Monastir through an improvement in the efficiency of stomatal conductance regulation. It also improved NaCl resistance in Sbikha through an increase in sodium excretion through the external bladders. It is suggested that polyamines (spermidine and spermine) are involved in the salt excretion process and that ABA contributes to polyamine synthesis as well as to the conversion from the bound and conjugated to the free soluble forms of polyamine. Proline accumulated in response to osmotic stress and slightly increased in response to ABA treatment while glycinebetaine accumulated in response to salinity and was not influenced by ABA. CONCLUSIONS: It is concluded that ABA is involved in both salt and osmotic stress resistance in the xerohalophyte species Atriplex halimus but that it acts on different physiological cues in response to those distinct environmental constraints.

Phytoextraction of Pb and Cd by the Mediterranean saltbush (Atriplex halimus L.): metal uptake in relation to salinity.

BACKGROUND, AIM, AND SCOPE: The success of phytoextraction depends upon the identification of suitable plant species that hyperaccumulate heavy metals and produce large amounts of biomass using established agricultural techniques. In this study, the Mediterranean saltbush Atriplex halimus L., which is a C4 perennial native shrub of Mediterranean basin with an excellent tolerance to drought and salinity, is investigated with the main aim to assess its phytoremediation potential for Pb and Cd removal from contaminated soils. In particular, the influence of soil salinity in metal accumulation has been studied as there is notable evidence that salinity changes the bioavailability of metals in soil and is a key factor in the translocation of metals from roots to the aerial parts of the plant. MATERIALS AND METHODS: Three pot experiments were conducted under greenhouse conditions for a 10-week period with A. halimus grown in soil artificially polluted with 20 ppm of Cd and/or 800 ppm of Pb and irrigated with three different salt solutions (0.0%, 0.5%, and 3.0% NaCl). Soil measurements for soil characterization were performed with the expiration of the first week of plant exposure to metals and NaCl, and at the end of the experimental period, chlorophyll content, leaf protein content, leaf specific activity of guaiacol peroxidase (EC 1.11.1.7), shoot water content, biomass, and Cd and Pb content in the plant tissues were determined. Additionally, any symptoms of metal or salt toxicity exhibited by the plants were visually noted during the whole experimental period. RESULTS: The experimental data suggest that increasing salinity increases cadmium uptake by A. halimus L. while in the case of lead there was not a clear effect of the presence of salt on lead accumulation in plant tissues. A. halimus developed no visible signs of metal toxicity; only salt toxicity symptoms were observed in plants irrigated with 3% NaCl solutions. Chlorophyll content, leaf protein content, shoot water content, and biomass were not negatively affected by the metals; instead, there was even an increase in the amount of photosynthetic pigments in plants treated with both metals and salinity. The specific activity of guaiacol peroxidase seems to have a general tendency for increase in plants treated with the metals in comparison with the respective controls but a statistically significant difference exists only in plants treated with the metal mixture and saline conditions. DISCUSSION: The data revealed that lead and cadmium accumulation in plant tissues was kept generally at low levels. Salinity was found to have a positive effect on cadmium uptake by the plant and this may be related to a higher bioavailability of the metal in soil due to decreased Cd sorption on soil particles. On the other hand, salinity did not influence in a clear way the uptake of Pb by the plant probably because of lead's limited mobility in soils and plant tissues. Cd and Pd usually decrease the chlorophyll content and biomass and change water relations in plants; however, A. halimus was found not to be affected indicating that it is a Cd- and Pb-tolerant plant. Guaiacol peroxidase activity as one of the parameters expressing oxidative damage and extent of stress in plants was not generally found to be significantly affected under the presence of metals in most plants suggesting that the extent of stress in plants was minimal, while only for plants treated with the metal mixture and low salinity the enzyme activity was elevated confirming that this enzyme serves as an antioxidative tool against the reactive oxygen species produced by the metals. CONCLUSIONS: Atriplex halimus L. is a Pb- and Cd-tolerant plant but metal concentrations achieved in plant tissues were kept generally at low levels; however, metal accumulation in shoots, especially for Cd, considered together with its high biomass production, rapid growth, and deep root system able to cope with poor structure and xeric characteristics of several polluted soils suggest that this plant deserves further investigation. RECOMMENDATIONS AND PERSPECTIVES: Phytoextraction by halophytes is a promising alternative for the remediation of heavy metal contaminated sites affected by salinity since saline depressions often indicate sites of industrial effluents accumulation, contaminated by heavy metals, including Pb and Cd. Halophytes are also promising candidates for the removal of heavy metals from non-saline soils. Furthermore, the use of such plants can be potentially viewed as an alternative method for soil desalination where salt is removed from the soil instead of being washed downwards by water or other solutions.

Response to non-uniform salinity in the root zone of the halophyte Atriplex nummularia: growth, photosynthesis, water relations and tissue ion concentrations.

BACKGROUND AND AIMS: Soil salinity is often heterogeneous, yet the physiology of halophytes has typically been studied with uniform salinity treatments. An evaluation was made of the growth, net photosynthesis, water use, water relations and tissue ions in the halophytic shrub Atriplex nummularia in response to non-uniform NaCl concentrations in a split-root system. METHODS: Atriplex nummularia was grown in a split-root system for 21 d, with either the same or two different NaCl concentrations (ranging from 10 to 670 mm), in aerated nutrient solution bathing each root half. KEY RESULTS: Non-uniform salinity, with high NaCl in one root half (up to 670 mm) and 10 mm in the other half, had no effect on shoot ethanol-insoluble dry mass, net photosynthesis or shoot pre-dawn water potential. In contrast, a modest effect occurred for leaf osmotic potential (up to 30 % more solutes compared with uniform 10 mm NaCl treatment). With non-uniform NaCl concentrations (10/670 mm), 90 % of water was absorbed from the low salinity side, and the reduction in water use from the high salinity side caused whole-plant water use to decrease by about 30 %; there was no compensatory water uptake from the low salinity side. Leaf Na(+) and Cl(-) concentrations were 1.9- to 2.3-fold higher in the uniform 670 mm treatment than in the 10/670 mm treatment, whereas leaf K(+) concentrations were 1.2- to 2.0-fold higher in the non-uniform treatment. CONCLUSIONS: Atriplex nummularia with one root half in 10 mm NaCl maintained net photosynthesis, shoot growth and shoot water potential even when the other root half was exposed to 670 mm NaCl, a concentration that inhibits growth by 65 % when uniform in the root zone. Given the likelihood of non-uniform salinity in many field situations, this situation would presumably benefit halophyte growth and physiology in saline environments.