Saltbush seedlings (Atriplex spp.) shed border-like cells from closed-type root apical meristems.

Australian saltbush (Atriplex spp.) survive in exceptionally saline environments and are often used for pasture in semi-arid areas. To investigate the impact of salinity on saltbush root morphology and root exudates, three Australian native saltbush species (Atriplex nummularia , Atriplex amnicola , and Atriplex vesicaria ) were grown in vitro in optimised sterile, semi-hydroponic systems in media supplemented with different concentrations of salt (NaCl). Histological stains and chromatographic techniques were used to characterise the root apical meristem (RAM) type and root exudate composition of the saltbush seedlings. We report that saltbush species have closed-type RAMs, which release border-like cells (BLCs). Monosaccharide content, including glucose and fructose, in the root mucilage of saltbush was found to be uniquely low, suggesting that saltbush may minimise carbon release in polysaccharides of root exudates. Root mucilage also contained notable levels of salt, plus increasing levels of unidentified compounds at peak salinity. Un-esterified homogalacturonan, xyloglucan, and arabinogalactan proteins between and on the surface of BLCs may aid intercellular adhesion. At the highest salinity levels, root cap morphology was altered but root:shoot ratio remained consistent. While questions remain about the identity of some components in saltbush root mucilage other than the key monosaccharides, this new information about root cap morphology and cell surface polysaccharides provides avenues for future research.

Comparative study of the anthelmintic activity of ethanolic and aqueous extracts of Atriplex halimus, the anti-echinococcal plant used in Algeria.

Cystic echinococcosis (CE) remains a major human public health problem in Algeria. Surgical treatment is the gold standard. However, one of the most critical complications of surgery is a recurrence of the disease, which is attributed to the dissemination of protoscoleces during the operation. This study aimed to compare the scolicidal effect of two ethanolic and aqueous extracts of Atriplex halimus, a plant used by the Algerian population against CE. The comparison involved the assessment of the phytochemical content, making screening using GC-MS/MS, and the evaluation of the antioxidant activity using DPPH and the in vitro scolicidal effect of A. halimus extracts. The aqueous and ethanolic extracts of leaves contained 19.601 ± 0.016 and 15.406 ± 0.003 mg of gallic acid equivalent per g of extract (GAE/g extract) of total phenolic. However, the flavonoid content in ethanolic and aqueous extracts was 4.350 ± 0.023 and 1.995 ± 0.026 mg of quercetin equivalent per g (QE/g extract), respectively. Four compounds were identified by GC-MS/MS for the ethanolic extract, while twenty-three compounds were determined for the aqueous extract. The ethanolic and aqueous extracts of A. halimus demonstrated an antioxidant activity with IC50 = 0.850 ± 0.026 mg/ml and IC50 = 0.897 ± 0.060 mg/ml, respectively. The higher in vitro scolicidal effect was 100% after 90 min and 10 min at 100 and 150 mg/ml, respectively for the ethanolic extract and after 120 and 60 min at 100 and 150 mg/ml, respectively for the aqueous extract. Atriplex halimus could be used as a natural source for the production of a potential scolicidal agent for use during CE surgery and/or percutaneous interventions.

Soil salinity regulates spatial-temporal heterogeneity of seed germination and seedbank persistence of an annual diaspore-trimorphic halophyte in northern China.

BACKGROUND AND AIMS: Seed heteromorphism is a plant strategy that an individual plant produces two or more distinct types of diaspores, which have diverse morphology, dispersal ability, ecological functions and different effects on plant life history traits. The aim of this study was to test the effects of seasonal soil salinity and burial depth on the dynamics of dormancy/germination and persistence/depletion of buried trimorphic diaspores of a desert annual halophyte Atriplex centralasiatica. METHODS: We investigated the effects of salinity and seasonal fluctuations of temperature on germination, recovery of germination and mortality of types A, B, C diaspores of A. centralasiatica in the laboratory and buried diaspores in situ at four soil salinities and three depths. Diaspores were collected monthly from the seedbank from December 2016 to November 2018, and the number of viable diaspores remaining (not depleted) and their germinability were determined. RESULTS: Non-dormant type A diaspores were depleted in the low salinity "window" in the first year. Dormant diaspore types B and C germinated to high percentages at 0.3 and 0.1 mol L-1 soil salinity, respectively. High salinity and shallow burial delayed depletion of diaspore types B and C. High salinity delayed depletion time of the three diaspore types and delayed dormancy release of types B and C diaspores from autumn to spring. Soil salinity modified the response of diaspores in the seedbank by delaying seed dormancy release in autum and winter and by providing a low-salt concentration window for germination of non-dormant diaspores in spring and early summer. CONCLUSIONS: Buried trimorphic diaspores of annual desert halophyte A. centralasiatica exhibited diverse dormancy/germination behavior in respond to seasonal soil salinity fluctuation. Prolonging persistence of the seedbank and delaying depletion of diaspores under salt stress in situ primarily is due to inhibition of dormancy-break. The differences in dormancy/germination and seed persistence in the soil seedbank may be a bet-hadging strategy adapted to stressful temporal and spatial heterogeneity, and allows A. centralasiatica to persist in the unpredictable cold desert enevironment.

Habitat-specific allocations of elements in Atriplex lentiformis seeds indicate adaptation to metal toxicity.

Self-sustaining vegetation in metal-contaminated areas is essential for rebuilding ecological resilience and community stability in degraded lands. Metal-tolerant plants originating from contaminated post-mining areas may hold the key to successful plant establishment and growth. Yet, little is known about the impact of metal toxicity on reproductive strategies, metal accumulation, and allocation patterns at the seed stage. Our research focused on the metal tolerant Atriplex lentiformis. Specifically, we examined the effects of toxic metal(loid) concentration in soils on variability in its reproductive strategies, including germination patterns, elemental uptake, and allocation within the seeds. We employed advanced imaging techniques like synchrotron X-ray fluorescence microscopy (2D scans and 3D tomograms) combined with inductively coupled plasma mass spectrometry to reveal significant differences in metal(loid) concentration and distribution within the seed structures of A. lentiformis from contrasting habitats. Exclusive Zn hotspots of high concentrations were found in the seeds of the metallicolous accession, primarily in the sensitive tissues of shoot apical meristems and root zones of the seed embryos. Our findings offer novel insights into phenotypic variability and metal tolerance and accumulation in plants from extreme environments. This knowledge can be applied to enhance plant survival and performance in land restoration efforts.

Drosophila melanogaster as a model organism for diabetes II treatment by the ethyl acetate fraction of Atriplex halimus L.

Type 2 diabetes (T2D) is the most common metabolic disorder. The undesirable effects of synthetic drugs demand a search for safe antidiabetic agents. This study aimed to assess the antidiabetic activity of different fractions of Atriplex halimus (petroleum ether 60-80, methylene chloride, ethyl acetate, and n-butanol) using Drosophila melanogaster larvae. Titers of total glucose and trehalose, as well as larval weight, were measured and compared with those of control and diabetic larvae. The expression of Drosophila insulin-like peptides (DILP2 and DILP3) and adipokinetic hormone (AKH) was evaluated. The results revealed a significant increase in total glucose, trehalose, and a decrease in body weight in the larvae fed a high-sugar diet compared with those in the control. When larvae fed diets containing the tested fractions, the total glucose and trehalose decreased to the control level, and the body weight increased. DILP2, DILP3, and AKH exhibited significant decreases upon treatment with A. halimus ethyl acetate. Metabolomic profiling of the ethyl acetate fraction of A. halimus revealed the presence of flavonoids and flavonoid glycosides. After docking screening to predict the most powerful moiety, we discovered that flavonoid glycosides (especially eriodictyol-7-O-neohesperidoside) have a greater affinity for the pocket than the other moieties. The results indicated the therapeutic activity of the A. halimus ethyl acetate fraction against induced T2D in Drosophila larvae. The antidiabetic activity may be attributed to flavonoids, which are the main components of the A. halimus ethyl acetate fraction.

Hortensins, Type 1 Ribosome-Inactivating Proteins from Seeds of Red Mountain Spinach: Isolation, Characterization, and Their Effect on Glioblastoma Cells.

Ribosome inactivating proteins (RIPs) are specific N-ß-glycosylases that are well-characterized in plants. Their enzymatic action is to damage ribosomes, thereby blocking protein translation. Recently, several research groups have been working on the screening for these toxins in edible plants to facilitate the use of RIPs as biotechnological tools and biopesticides and to overcome public prejudice. Here, four novel monomeric (type 1) RIPs have been isolated from the seeds of Atriplex hortensis L. var. rubra, which is commonly known as edible red mountain spinach. These enzymes, named hortensins 1, 2, 4, and 5, are able to release the ß-fragment and, like many other RIPs, adenines from salmon sperm DNA, thus, acting as polynucleotide:adenosine glycosidases. Structurally, hortensins have a different molecular weight and are purified with different yields (hortensin 1, ~29.5 kDa, 0.28 mg per 100 g; hortensin 2, ~29 kDa, 0.29 mg per 100 g; hortensin 4, ~28.5 kDa, 0.71 mg per 100 g; and hortensin 5, ~30 kDa, 0.65 mg per 100 g); only hortensins 2 and 4 are glycosylated. Furthermore, the major isoforms (hortensins 4 and 5) are cytotoxic toward human continuous glioblastoma U87MG cell line. In addition, the morphological change in U87MG cells in the presence of these toxins is indicative of cell death triggered by the apoptotic pathway, as revealed by nuclear DNA fragmentation (TUNEL assay).

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 Analysis, Acetylcholinesterase Inhibition, Antidiabetic and Antioxidant Activities of Atriplex halimus L. (Amaranthaceae Juss.).

Mediterranean saltbush Atriplex halimus L. (Amaranthaceae) from different bioclimatic arid zones (ten wild populations) were studied. Phenols contents, flavonoids, flavonols, tannins and anthocyanins were determined and then tested for their antioxidants, antidiabetic and anti-acetylcholinesterase (AChE) activities. Levels of total polyphenols including flavonoids and flavonols, tannins and anthocyanins were high and varied significantly among analyzed populations. Nine phenolic acids and four flavonoids were identified for the first time in the methanolic fraction and quantified by liquid high-performance chromatography system HPLC (DAD). All extracts showed a substantial antioxidant activity, as assessed by DPPH assay (1,1-diphenyl-2-picrylhydrazyl free radical) (IC50DPPH=147.3for population of Seliena), Ferric Reducing Antioxidant Power (FRAP; IC50FRAP=3.2 for populations of Sousse and Kairouan), and Chelation Fer test (IC50FerCh=1.5 µg/mL for populations of El-hamma and Mednine). Atriplex halimus possessed a high inhibitory effect against α-amylase activity (up to 2.6 mg ACE/gE), a moderate activity for α-glucosidase (up to 91.0 mg ACE/gE) and AChE (up to 147.2 µg/mL) compared to standard. The analyzed populations were isolated and subdivided into three distinct groups, without any bioclimatic structuration. Enzymatic activities seem to be associated with the presence, in plant extracts, of other classes of compounds then phenols such as terpenes, sterols, saponins, coumarins and carotenoids.

Phytochemical characterization, in vivo and in vitro biological activities of Atriplex lindleyi Moq. subsp. inflata.

ETHNOPHARMACOLOGICAL RELEVANCE: Atriplex lindleyi Moq. subsp. inflata from Algeria is known for its traditional use for therapeutic properties. AIM OF THE STUDY: We analysed quantitatively and qualitatively the infusion and diethyl ether, n-butanol and ethereal extracts of A. lindleyi, as well as assessed their acute toxicity, anti-inflammatory, antinociceptive and antioxidant activities. MATERIALS AND METHODS: The total phenolic and flavonoid contents were quantified. The anti-inflammatory effect was assessed using a carrageenan-induced paw oedema assay and the antinociceptive effect was evaluated using an acetic acid-induced writhing method. In addition, antioxidant activity was examined by three tests: DPPH, reducing power and phenanthroline assays. RESULTS: The quantity of total phenols in the roots of A. lindleyi was larger than in the aerial parts, while the amounts of total flavonoids were larger in the aerial parts than in the roots. The HPLC profiles allowed us to identify 32 compounds belonging to the phenolic acid and flavonoid classes. Intraperitoneal administration of the infusions and phenolic extracts in mice did not cause any symptoms of toxicity or mortality. Results revealed that the aerial parts and roots of A. lindleyi had potential anti-inflammatory and antinociceptive activities and were higher than diclofenac and paracetamol, respectively. The diethyl ether extract (DEE) of aerial parts and roots showed the greatest antioxidant activity in comparison to the other tested extracts. CONCLUSION: The phenolic composition attempted to be identified by HPLC confers this plant's pharmacological potential. The antioxidant potential may be due to active ingredients, including ascorbic acid, in the two studied parts of A. lindleyi, which is a bioactive molecule with strong antioxidant properties. Also, detecting salicylic acid, the active antinociceptive ingredient of aspirin, gallic and ferulic acids may justify this subspecies' antinociceptive and anti-inflammatory potentials.

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.

Flavonoids are involved in salt tolerance through ROS scavenging in the halophyte Atriplex canescens.

KEY MESSAGE: The content of flavonoids could increase in A. canescens under saline conditions. Overexpression of AcCHI in transgenic A. thaliana promotes flavonoid biosynthesis, thereby functioning in the tolerance of transgenic plants to salt and osmotic stress by maintaining ROS homeostasis. Atriplex canescens is a halophytic forage shrub with excellent adaptation to saline environment. Our previous study showed that a large number of genes related to the biosynthesis of flavonoids in A. canescens were significantly up-regulated by NaCl treatments. However, it remains unclear whether flavonoids are involved in A. canescens response to salinity. In this study, we found that the accumulation of flavonoids significantly increased in either the leaves or roots of A. canescens seedling under 100 and 300 mM NaCl treatments. Correspondingly, AcCHS, AcCHI and AcF3H, which encode three key enzymes (chalcone synthases (CHS), chalcone isomerase (CHI), and flavanone 3-hydroxylase (F3H), respectively) of flavonoids biosynthesis, were significantly induced in the roots or leaves of A. canescens by 100 or 300 mM NaCl. Then, we generated the transgenic Arabidopsis thaliana overexpressing AcCHI and found that transgenic plants accumulated more flavonoids through enhancing the pathway of flavonoids biosynthesis. Furthermore, overexpression of AcCHI conferred salt and osmotic stress tolerance in transgenic A. thaliana. Contrasted with wild-type A. thaliana, transgenic lines grew better with greater biomass, less H2O2 content as well as lower relative plasma permeability in either salt or osmotic stress conditions. In conclusion, our results indicate that flavonoids play an important role in A. canescens response to salt stress through reactive oxygen species (ROS) scavenging and the key enzyme gene AcCHI in flavonoids biosynthesis pathway of A. canescens has the potential to improve the stress tolerance of forages and crops.

Unveiling Alternative Oxidation Pathways and Antioxidant and Cardioprotective Potential of Amaranthin-Type Betacyanins from Spinach-like Atriplex hortensis var. 'Rubra'.

A comprehensive oxidation mechanism was investigated for amaranthin-type betacyanins with a specific glucuronosylglucosyl moiety isolated from Atriplex hortensis 'rubra' using liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS) and LC-Quadrupole-Orbitrap-MS (LC-Q-Orbitrap-MS). By employing one-dimensional (1D) and two-dimensional (2D) NMR, this study elucidates the chemical structures of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS)-oxidized celosianins for the first time. These findings demonstrate alternative oxidation pathways for acylated betacyanins compared to well-known betanidin, betanin, and gomphrenin pigments. Contrary to previous research, we uncover the existence of 17-decarboxy-neo- and 2,17-bidecarboxy-xanneo-derivatives as the initial oxidation products without the expected 2-decarboxy-xan forms. These oxidized compounds demonstrated potent free radical scavenging properties. Celosianin (IC50 = 23 µg/mL) displayed slightly higher antioxidant activity compared to oxidized forms, 17-decarboxy-neocelosianin (IC50 = 34 µg/mL) and 2,17-bidecarboxy-xanneocelosianin (IC50 = 29 µg/mL). The oxidized compounds showed no cytotoxic effects on H9c2 rat cardiomyoblasts (0.1-100 µg/mL). Additionally, treatment of H9c2 cells with the oxidized compounds (0.1-10 µg/mL) elevated glutathione levels and exhibited protective effects against H2O2-induced cell death. These findings have significant implications for understanding the impact of oxidation processes on the structures and biological activities of acylated betalains, providing valuable insights for future studies of the bioavailability and biological mechanism of their action in vivo.

Screening of tolerance of Atriplex vulgatissima under zinc or lead experimental conditions. An integrative perspective by using the integrated biological response index (IBRv2).

The search for plants with a high capacity to tolerate and accumulate metals is an important issue in phytoremediation. In this sense, this study was conducted in the halophyte Atriplex vulgatissima to evaluate the effects of different concentrations of lead (Pb, 50 and 100 µM) or zinc (Zn, 100 and 200 µM) on morphological, physiological, and biochemical parameters as well as the accumulation patterns of this species. The results indicated that while essential metal Zn showed high translocation from roots to shoots (TF > 1), non-essential Pb was mainly accumulated in the roots (BCF>1). Regarding shape, both metals induced slenderness of the blade, but only Zn treatment reduced leaf size. No difference in biomass production and photosynthetic parameters was found between Pb and Zn treatments. Pb treatments did not show significant differences between treatments regarding water content (WC), pigment concentration, and the activity of superoxide dismutase (SOD) and guaiacol peroxidase (GPx), but did result in a decrease in catalase activity at 100 µM Pb. On the other hand, 200 µM Zn leads to a clear reduction in WC and pigment concentrations, along with an increase in SOD and GPx activities. In addition, ascorbate peroxidase (APx) activity showed a hormesis effect at 50 µM Pb and 100 µM Zn. Malondialdehyde increased with both Pb and Zn treatments. The integrated biological index (IBRv2) indicated that 200 µM Zn was the most affected treatment (IBRv2 = 19.02) and that under the same concentrations of metals (100 µM Pb or Zn), Pb treatments presented major stress (IBRv2 = 11.55). A. vulgatissima is a metallophyte with the potential for Pb phytostabilization and Zn phytoextraction, as well as a bioindicator of these metals. Its high biomass and deep roots, combined with its halophytic traits, make it suitable for bioremediation and monitoring programs.

The nutritional composition of six plant species after irrigation with treated wastewater and possible hazards by heavy metal accumulation.

A field experiment was conducted investigating the possibility of using treated wastewater (TWW) on sites affected by water scarcity in summer, waterlogging during the wet season, and salinity. A corresponding pot experiment was conducted comparable to the field experiment in Kalaât Landelous. The same plant species (Atriplex nummularia Lindl., Eucalyptus gomphocephala DC., Acacia cyanophylla Lindl., Casuarina glauca Sieber ex Spreng., Cupressus sempervirens L., and Pinus halepensis Mill.) were grown with the same treatments. While, in the field the plants, elemental composition cannot be linked to inputs by TWW, this was studied under controlled conditions. Additionally, a control was established lower in salinity receiving tap water. The effect of TWW irrigation on macro- and microelement uptake by the six plant species was studied. The treatments were high soil salinity under drained saline (DS) conditions, high salinity under waterlogged saline (WS), and a drained non-saline control (DNS: EC = 3.0 dS/m, pH = 8.4). TWW application under DS treatment increased Na, Cl, Ca, Mg, N, P, and K in most plant tissues compared to the control. TWW application in WS treatment resulted in an increase in heavy metals. Cu and Zn showed the highest bioaccumulation factor (BAF). The BAF in different plant tissues followed the order: Cu > Zn > Mn > Cd > Ni > Co > Pb. The plants accumulated significant amounts of metals in their roots.

Zinc accumulation in Atriplex lentiformis is driven by plant genes and the soil microbiome.

Successful phytoremediation of acidic metal-contaminated mine tailings requires amendments to condition tailings properties prior to plant establishment. This conditioning process is complex and includes multiple changes in tailings bio-physico-chemical properties. The objective of this project is to identify relationships between tailings properties, the soil microbiome, and plant stress response genes during growth of Atriplex lentiformis in compost-amended (10 %, 15 %, 20 % w/w) mine tailings. Analyses include RNA-Seq for plant root gene expression, 16S rRNA amplicon sequencing for bacterial/archaeal communities, metal concentrations in both tailings and plant organs, and phenotypic measures of plant stress. Zn accumulation in A. lentiformis leaves varied with compost levels and was the highest in the intermediate treatment (15 %, TC15). Microbial analysis identified Alicyclobacillus, Hydrotalea, and Pseudolabrys taxa with the highest relative abundance in TC15, and these taxa were strongly associated with Zn accumulation. Furthermore, we identified 190 root genes with significant gene expression changes. These root genes were associated with different pathways including, abscisic acid and auxin signaling, defense responses, ion channels, metal ion binding, oxidative stress, transcription regulation, and transmembrane transport. However, root gene expression changes were not driven by the increasing levels of compost. For example, there were 15 genes that were up-regulated in TC15, whereas 106 genes were down-regulated in TC15. The variables analyzed explained 86 % of the variance in Zn accumulation in A. lentiformis leaves. Importantly, Zn accumulation was driven by Zn shoot concentrations, leaf stress symptoms, plant root genes, and microbial taxa. Therefore, our results suggest there are strong plant-microbiome associations that drive Zn accumulation in A. lentiformis and different plant gene pathways are involved in alleviating varying levels of metal stress. Future work is needed to gain a mechanistic understanding of these plant-microbiome interactions to optimize phytoremediation strategies as they will govern the success or failure of the revegetation process.

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.

Effect of jasmonic acid on the phytoremediation of dinitrophenol from wastewater by Solanum nigrum L. and Atriplex lentiformis (Torr.) S. Watson.

Phytoremediation is one of the best methods for cleaning up natural resources like water because plants are eco-friendly and safe for the ecosystem. Hyperaccumulators, e.g., Solanum nigrum L. and Atriplex lentiformis (Torr.) S. Watson, have been used to remove toxic metals from soil and water through phytoremediation techniques, but it is unknown if they can remove hazardous chemicals such as dinitrophenol (DNP), from wastewater. A hydroponic experiment was conducted to study the efficiency of S. nigrum and A. lentiformis in removing DNP from wastewater. Jasmonic acid (JAC) was applied to the tested plants in two doses, 0.25 and 0.50 mmol, in an effort to better understand how it affects phytoremediation effectiveness. The growth of S. nigrum and A. lentiformis improved significantly (p < 0.05) by the foliar application of JAC. The applications of JAC1 and JAC2 significantly (p < 0.05) increased nutrient uptake and chlorophyll concentrations in S. nigrum and A. lentiformis plants. The foliar spraying of S. nigrum and A. lentiformis with JAC significantly (p < 0.05) increased the antioxidant enzymes activity, i.e., SOD and POD. The levels of osmoregulatory substances like proline and carbohydrates significantly (p < 0.05) increased after JAC was sprayed on S. nigrum and A. lentiformis plants. In the case of S. nigrum, the efficiency of DNP removal varied between 53 and 69%, with an average of 63%, while in the case of A. lentiformis, it varied between 47 and 62%, with an average of 56%. The removal efficiency of DNP reached 67 and 69% when S. nigrum was sprayed with JAC1 and JAC2. When JAC1 and JAC2 were sprayed on A. lentiformis, DNP removal efficiency rose from 47 to 60 and from 47 to 62%, respectively. S. nigrum and A. lentiformis plants can be grown normally and survive in dinitrophenol-contaminated water without showing any toxic symptoms. S. nigrum and A. lentiformis have a powerful antioxidant system and the ability to produce vital compounds that alleviate the stress caused by DNP toxicity. The findings are crucial for cleaning up polluted water and protecting the ecosystem's health from dangerous pollutants.

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.

Possible accumulation of critical metals in plants that hyperaccumulate their chemical analogues?

Lithium (Li), gallium (Ga) and indium (In) are industry-critical metals, with no known plant species that (hyper)accumulate these metals to any substantial degree. We hypothesised that sodium (Na) hyperaccumulators (i.e., halophytes) may accumulate Li, whilst aluminium (Al) hyperaccumulators may accumulate Ga and In, based on the chemical similarities of these elements. Experiments were conducted in hydroponics at various molar ratios for six weeks to determine accumulation in roots and shoots of the target elements. For the Li experiment, the halophytes Atriplex amnicola, Salsola australis and Tecticornia pergranulata were subjected to Na and Li treatments, whilst for the Ga and In experiment, Camellia sinensis was exposed to Al, Ga, and In. The halophytes were able to accumulate high shoot Li and Na concentrations reaching up to ~10 g Li kg-1 and 80 g Na kg-1, respectively. The translocation factors for Li were higher than for Na (about two-fold) in A. amnicola and S. australis. The results from the Ga and In experiment show that C. sinensis is capable of accumulating high concentrations of Ga (mean 150 mg Ga kg-1), comparable with Al (mean 300 mg Al kg-1), but virtually no In (<20 mg In kg-1) in its leaves. Competition between Al and Ga suggests that Ga might be taken up via Al pathways in C. sinensis. The findings suggest that there are opportunities to explore Li and Ga phytomining on respective Li- and Ga-enriched mine water/soil/mine waste materials using halophytes and Al hyperaccumulators to complement the global supply of these critical metals.

Phytoremediation of dinitrophenol from wastewater by atriplex lentiformis: effect of salicylic acid.

Quail bush [Atriplex lentiformis (Torr.) S. Wats] plants were used in removing 2, 4-dinitrophenol (DNP) from wastewater in a hydroponic experiment. The hydroponic system contained three doses of DNP, i.e., 0, 10, and 20 mg L-1. Quail bush plants were sprayed with 0.1 mM salicylic acid (SA) to study its role in resisting DNP toxicity. DNP significantly (p < 0.05) reduced plant growth. Exposure of A. lentiformis plants to 20 mg L-1 of DNP reduced the total chlorophyl and relative water content by 39 and 24%, respectively. SA improved the antioxidant defense in terms of ascorbate peroxidase (APX) and polyphenol oxidase (PPO) activities. SA alleviated DNP toxicity by enhancing the production of osmoprotectants, e.g.,proline, phenols, and carbohydrates. SA enhanced the removal efficiency of DNP and the highest removal efficiency (96%) was recorded in the plants sprayed with SA and grown on 10 mg L-1 of DNP. A. lentiformis is a halophytic plant that has good physiological characteristics to resist 2, 4-dinitrophenol toxicity in wastewaters and is qualified to purify water from these harmful compounds. Exogenous application of 0.1 mM SA increased the defense system in A. lentiformis against 2, 4-dinitrophenol toxicity and enhanced the removal efficiency.

2, 4-dinitrophenol inhibited the synthesis of photosynthetic pigments.Salicylic acid protects the vital bio-compounds in plant cells.Atriplex plants are able to remove (96%) of 2, 4-dinitrophenol from the wastewater.Atriplex plants have a strong antioxidant defense enable them to survive in wastewater.