Unravelling the halophyte Suaeda maritima as an efficient candidate for phytostabilization of cadmium and lead: Implications from physiological, ionomic, and metabolomic responses.

Cadmium (Cd) and lead (Pb) are among the most toxic heavy metals affecting human health and crop yield. Suaeda maritima (L.) Dumort is an obligate halophyte that is well adapted to saline soil. The inbuilt salinity tolerance mechanisms of halophytes help them to survive in heavy metal-contaminated rhizospheric soil. In the present study, growth and ionomic responses, reactive oxygen species (ROS) accumulation, modulations of phytochelatins, antioxidative defense, and metabolomic responses were studied in S. maritima imposed to Cd and Pb stresses with an aim to elucidate Cd and Pb tolerance mechanisms and phytoremediation potential of this halophyte. Our results showed a reduction of biomass in S. maritima, which may serve as an energy conservation strategy for survival under heavy metal stress. The increased accumulation of ROS with concomitant higher expression of various antioxidative enzymes suggests the efficient scavenging of ROS. The metabolite profiling revealed significant up-regulation of sugars, sugar alcohols, amino acids, polyphenols, and organic acids under Cd and Pb stresses suggesting their possible role in osmotic balance, ionic homeostasis, ROS scavenging, and signal transduction for stress tolerance. In S. maritima, the translocation factors (Tf) are <1 in both Cd and Pb treatments, which indicates that this halophyte has high phytostabilization potential for Cd and Pb in roots and through restricted translocation of heavy metal ions to the aboveground part. The findings of this study offer comprehensive information on Cd and Pb tolerance mechanisms in S. maritima and suggest that this halophyte can detoxify the HMs through physiological, ionic, antioxidative, and metabolic regulations.

Modulation of physio-biochemical and photosynthesis parameters by overexpressing SbPIP2 gene improved abiotic stress tolerance of transgenic tobacco.

The present study aims to explore the potential of a plasma-membrane localized PIP2-type aquaporin protein sourced from the halophyte Salicornia brachiata to alleviate salinity and water deficit stress tolerance in a model plant through transgenic intervention. Transgenic plants overexpressing SbPIP2 gene showed improved physio-biochemical parameters like increased osmolytes (proline, total sugar, and amino acids), antioxidants (polyphenols), pigments and membrane stability under salinity and drought stresses compared to control plants [wild type (WT) and vector control (VC) plants]. Multivariate statistical analysis showed that, under water and salinity stresses, osmolytes, antioxidants and pigments were correlated with SbPIP2-overexpressing (SbPIP2-OE) plants treated with salinity and water deficit stress, suggesting their involvement in stress tolerance. As aquaporins are also involved in CO2 transport, SbPIP2-OE plants showed enhanced photosynthesis performance than wild type upon salinity and drought stresses. Photosynthetic gas exchange (net CO2 assimilation rate, PSII efficiency, ETR, and non-photochemical quenching) were significantly higher in SbPIP2-OE plants compared to control plants (wild type and vector control plants) under both unstressed and stressed conditions. The higher quantum yield for reduction of end electron acceptors at the PSI acceptor side [Φ( R0 )] in SbPIP2-OE plants compared to control plants under abiotic stresses indicates a continued PSI functioning, leading to retained electron transport rate, higher carbon assimilation, and less ROS-mediated injuries. In conclusion, the SbPIP2 gene functionally validated in the present study could be a potential candidate for engineering abiotic stress resilience in important crops.

Novel Proteins of the High-Affinity Nitrate Transporter Family NRT2, SaNRT2.1 and SaNRT2.5, from the Euhalophyte Suaeda altissima: Molecular Cloning and Expression Analysis.

Two genes of nitrate transporters SaNRT2.1 and SaNRT2.5, putative orthologs of high-affinity nitrate transporter genes AtNRT2.1 and AtNRT2.5 from Arabidopsis thaliana, were cloned from the euhalophyte Suaeda altissima. Phylogenetic bioinformatic analysis demonstrated that the proteins SaNRT2.1 and SaNRT2.5 exhibited higher levels of homology to the corresponding proteins from the plants of family Amaranthaceae; the similarity of amino acid sequences between proteins SaNRT2.1 and SaNRT2.5 was lower (54%). Both SaNRT2.1 and SaNRT2.5 are integral membrane proteins forming 12 transmembrane helices as predicted by topological modeling. An attempt to demonstrate nitrate transporting activity of SaNRT2.1 or SaNRT2.5 by heterologous expression of the genes in the yeast Hansenula (Ogataea) polymorpha mutant strain Δynt1 lacking the only yeast nitrate transporter was not successful. The expression patterns of SaNRT2.1 and SaNRT2.5 were studied in S. altissima plants that were grown in hydroponics under either low (0.5 mM) or high (15 mM) nitrate and salinity from 0 to 750 mM NaCl. The growth of the plants was strongly inhibited by low nitrogen supply while stimulated by NaCl; it peaked at 250 mM NaCl for high nitrate and at 500 mM NaCl for low nitrate. Under low nitrate supply, nitrate contents in S. altissima roots, leaves and stems were reduced but increased in leaves and stems as salinity in the medium increased. Potassium contents remained stable under salinity treatment from 250 to 750 mM NaCl. Quantitative real-time PCR demonstrated that without salinity, SaNRT2.1 was expressed in all organs, its expression was not influenced by nitrate supply, while SaNRT2.5 was expressed exclusively in roots-its expression rose about 10-fold under low nitrate. Salinity increased expression of both SaNRT2.1 and SaNRT2.5 under low nitrate. SaNRT2.1 peaked in roots at 500 mM NaCl with 15-fold increase; SaNRT2.5 peaked in roots at 500 mM NaCl with 150-fold increase. It is suggested that SaNRT2.5 ensures effective nitrate uptake by roots and functions as an essential high-affinity nitrate transporter to support growth of adult S. altissima plants under nitrogen deficiency.

Extraction of cellulose from halophytic plants for the synthesis of a novel biocomposite.

Cellulose nanofibers, a sustainable and promising material with widespread applications, exhibit appreciable strength and excellent mechanical and physicochemical properties. The preparation of cellulosic nanofibers from food or agricultural residue is not sustainable. Therefore, this study was designed to use three halophytic plants (Cressa cretica, Phragmites karka, and Suaeda fruticosa) to extract cellulose for the subsequent conversion to cellulosic nanofibers composites. The other extracted biomass components including lignin, hemicellulose, and pectin were also utilized to obtain industrially valuable enzymes. The maximum pectinase (31.56 IU mL-1), xylanase (35.21 IU mL-1), and laccase (15.89 IU mL-1) were produced after the fermentation of extracted pectin, hemicellulose, and lignin from S. fruticosa, P. karka, and C. cretica, respectively. Cellulose was methylated (with a degree of substitution of 2.4) and subsequently converted into a composite using polyvinyl alcohol. Scanning electron microscopy and Fourier-transform infrared spectroscopy confirmed the successful synthesis of the composites. The composites made up of cellulose from C. cretica and S. fruticosa had a high tensile strength (21.5 and 15.2 MPa) and low biodegradability (47.58% and 44.56%, respectively) after dumping for 3 months in soil, as compared with the composite from P. karka (98.79% biodegradability and 4.9 MPa tensile strength). Moreover, all the composites exhibited antibacterial activity against gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae) and gram-positive bacteria (Staphylococcus aureus). Hence, this study emphasizes the possibility for various industrial applications of biomass from halophytic plants.

SOS1 tonoplast neo-localization and the RGG protein SALTY are important in the extreme salinity tolerance of Salicornia bigelovii.

The identification of genes involved in salinity tolerance has primarily focused on model plants and crops. However, plants naturally adapted to highly saline environments offer valuable insights into tolerance to extreme salinity. Salicornia plants grow in coastal salt marshes, stimulated by NaCl. To understand this tolerance, we generated genome sequences of two Salicornia species and analyzed the transcriptomic and proteomic responses of Salicornia bigelovii to NaCl. Subcellular membrane proteomes reveal that SbiSOS1, a homolog of the well-known SALT-OVERLY-SENSITIVE 1 (SOS1) protein, appears to localize to the tonoplast, consistent with subcellular localization assays in tobacco. This neo-localized protein can pump Na+ into the vacuole, preventing toxicity in the cytosol. We further identify 11 proteins of interest, of which SbiSALTY, substantially improves yeast growth on saline media. Structural characterization using NMR identified it as an intrinsically disordered protein, localizing to the endoplasmic reticulum in planta, where it can interact with ribosomes and RNA, stabilizing or protecting them during salt stress.

Ectopic expression of HaPEPC1 from the desert shrub Haloxylon ammodendron confers drought stress tolerance in Arabidopsis thaliana.

Phosphoenolpyruvate carboxylase (PEPC) plays a crucial role in the initial carbon fixation process in C4 plants. However, its nonphotosynthetic functions in Haloxylon ammodendron, a C4 perennial xerohalophytic shrub, are still poorly understood. Previous studies have reported the involvement of PEPC in plant responses to abiotic stresses such as drought and salt stress. However, the underlying mechanism of PEPC tolerance to drought stress has not been determined. In this study, we cloned the C4-type PEPC gene HaPEPC1 from H. ammodendron and investigated its biological function by generating transgenic Arabidopsis plants with ectopic expression of HaPEPC1. Our results showed that, compared with WT (wild-type) plants, ectopic expression of HaPEPC1 plants exhibited significantly greater germination rates and chlorophyll contents. Furthermore, under drought stress, the transgenic plants presented increased root length, fresh weight, photosynthetic capacity, and antioxidant enzyme activities, particularly ascorbate peroxidase and peroxidase. Additionally, the transgenic plants exhibited reduced levels of malondialdehyde, H2O2 (hydrogen peroxide), and O2- (superoxide radical). Transcriptome analysis indicated that ectopic expression of HaPEPC1 primarily regulated the expression of genes associated with the stress defence response, glutathione metabolism, and abscisic acid (ABA) synthesis and signalling pathways in response to drought stress. Taken together, these findings suggest that the ectopic expression of HaPEPC1 enhances the reduction of H2O2 and O2- in transgenic plants, thereby improving reactive oxygen species (ROS) scavenging capacity and enhancing drought tolerance. Therefore, the HaPEPC1 gene holds promise as a candidate gene for crop selection aimed at enhancing drought tolerance.

Introducing the halophyte Salicornia europaea to investigate combined impact of salt and tidal submergence conditions.

Tolerance mechanisms to single abiotic stress events are being investigated in different plant species, but how plants deal with multiple stress factors occurring simultaneously is still poorly understood. Here, we introduce Salicornia europaea as a species with an extraordinary tolerance level to both flooding and high salt concentrations. Plants exposed to 0.5MNaCl (mimicking sea water concentrations) grew larger than plants not exposed to salt. Adding more salt reduced growth, but concentrations up to 2.5MNaCl were not lethal. Regular tidal flooding with salt water (0.5MNaCl) did not affect growth or chlorophyll fluorescence, whereas continuous flooding stopped growth while plants survived. Quantitative polymerase chain reaction (qPCR) analysis of plants exposed to 1% oxygen in air revealed induction of selected hypoxia responsive genes, but these genes were not induced during tidal flooding, suggesting that S. europaea did not experience hypoxic stress. Indeed, plants were able to transport oxygen into waterlogged soil. Interestingly, sequential exposure to salt and hypoxic air changed the expression of several but not all genes as compared to their expression upon hypoxia only, demonstrating the potential to use S . europaea to investigate signalling-crosstalk between tolerance reactions to multiple environmental perturbations.

Designing a T-cell epitope-based vaccine using in silico approaches against the Sal k 1 allergen of Salsola kali plant.

Allergens originated from Salsola kali (Russian thistle) pollen grains are one of the most important sources of aeroallergens causing pollinosis in desert and semi-desert regions. T-cell epitope-based vaccines (TEV) are more effective among different therapeutic approaches developed to alleviate allergic diseases. The physicochemical properties, and B as well as T cell epitopes of Sal k 1 (a major allergen of S. kali) were predicted using immunoinformatic tools. A TEV was constructed using the linkers EAAAK, GPGPG and the most suitable CD4+ T cell epitopes. RS04 adjuvant was added as a TLR4 agonist to the amino (N) and carboxyl (C) terminus of the TEV protein. The secondary and tertiary structures, solubility, allergenicity, toxicity, stability, physicochemical properties, docking with immune receptors, BLASTp against the human and microbiota proteomes, and in silico cloning of the designed TEV were assessed using immunoinformatic analyses. Two CD4+ T cell epitopes of Sal k1 that had high affinity with different alleles of MHC-II were selected and used in the TEV. The molecular docking of the TEV with HLADRB1, and TLR4 showed TEV strong interactions and stable binding pose to these receptors. Moreover, the codon optimized TEV sequence was cloned between NcoI and XhoI restriction sites of pET-28a(+) expression plasmid. The designed TEV can be used as a promising candidate in allergen-specific immunotherapy against S. kali. Nonetheless, effectiveness of this vaccine should be validated through immunological bioassays.

The novel chaperonin 10 like protein (SbCPN10L) from Salicornia brachiata (Roxb.) augment the heat stress tolerance in transgenic tobacco.

The heat stress is a significant environmental challenge and impede the plant growth, development and productivity. The characterization and utilization of novel genes for improving stress tolerance represents a paramount approach in crop breeding. In the present study, we report on cloning of a novel heat-induced chaperonin 10-like gene (SbCPN10L) from Salicornia brachiata and elucidation of its in-planta role in conferring the heat stress endurance. The transgenic tobacco over-expressing SbCPN10L gene exhibited enhanced growth attributes such as higher rate of seed germination, germination and vigor index at elevated (35 ± 1 °C) temperature (eT). The SbCPN10L tobacco exhibited greenish and healthy seedling growth under stress. Compared with control tobacco at eT, the transgenic tobacco had higher water contents, membrane stability index, stress tolerance index and photosynthetic pigments. Lower electrolyte leakage and less accumulation of malondialdehyde, hydrogen peroxide and reactive oxygen species indicated better heat stress tolerance in transgenic tobacco over-expressing SbCPN10L gene. Transgenic tobacco accumulated higher contents of sugars, starch, amino acids and polyphenols at eT. The negative solute potential observed in transgenic tobacco contributed to maintain water content and support improved growth under stress. The up-regulation of NtAPX, NtPOX and NtSOD in transgenic tobacco under stress indicated higher ROS scavenging ability and better physiological conditioning. The results recommend the SbCPN10L gene as a potential candidate gene with an ability to confer heat stress tolerance for climate resilient crops.

Metabolomic analysis reveals key metabolites and metabolic pathways in Suaeda salsa under salt and drought stress.

Suaeda salsa is an important salt- and drought-tolerant plant with important ecological restoration roles. However, little is known about its underlying molecular regulatory mechanisms. Therefore, understanding the response mechanisms of plants to salt and drought stress is of great importance. In this study, metabolomics analysis was performed to evaluate the effects of salt and drought stress on S. salsa . The experiment consisted of three treatments: (1) control (CK); (2) salt stress (Ps); and (3) drought stress (Pd). The results showed that compared with the control group, S. salsa showed significant differences in phenotypes under salt and drought stress conditions. First, a total of 207 and 292 differential metabolites were identified in the Ps/CK and Pd/CK groups, respectively. Second, some soluble sugars and amino acids, such as raffinose, maltopentoses, D -altro-beptulose, D -proline, valine-proline, proline, tryptophan and glycine-L -leucine, showed increased activity under salt and drought stress conditions, suggesting that these metabolites may be responsible for salt and drought resistance in S. salsa . Third, the flavonoid biosynthetic and phenylalanine metabolic pathways were significantly enriched under both salt and drought stress conditions, indicating that these two metabolic pathways play important roles in salt and drought stress resistance in S. salsa . The findings of this study provide new insights into the salt and drought tolerance mechanisms of S. salsa .

Molecular regulation of lipid metabolism in Suaeda salsa.

Suaeda salsa is remarkable for its high oil content and abundant unsaturated fatty acids. In this study, the regulatory networks on fatty acid and lipid metabolism were constructed by combining the de novo transcriptome and lipidome data. Differentially expressed genes (DEGs) associated with lipids biosynthesis pathways were identified in the S. salsa transcriptome. DEGs involved in fatty acid and glycerolipids were generally up-regulated in leaf tissues. DEGs for TAG assembly were enriched in developing seeds, while DEGs in phospholipid metabolic pathways were enriched in root tissues. Polar lipids were extracted from S. salsa tissues and analyzed by lipidomics. The proportion of galactolipid MGDG was the highest in S. salsa leaves. The molar percentage of PG was high in the developing seeds, and the other main phospholipids had higher molar percentage in roots of S. salsa. The predominant C36:6 molecular species indicates that S. salsa is a typical 18:3 plant. The combined transcriptomic and lipidomic data revealed that different tissues of S. salsa were featured with DEGs associated with specific lipid metabolic pathways, therefore, represented unique lipid profiles. This study will be helpful on understanding lipid metabolism pathway and exploring the key genes involved in lipid synthesis in S. salsa.

Ectopic over-expression of HaFT-1, a 14-3-3 protein from Haloxylon ammodendron, enhances acquired thermotolerance in transgenic Arabidopsis.

Haloxylon ammodendron, an important shrub utilized for afforestation in desert areas, can withstand harsh ecological conditions such as drought, high salt and extreme heat. A better understanding of the stress adaptation mechanisms of H. ammodendron is vital for ecological improvement in desert areas. In this study, the role of the H. ammodendron 14-3-3 protein HaFT-1 in thermotolerance was investigated. qRT-PCR analysis showed that heat stress (HS) priming (the first HS) enhanced the expression of HaFT-1 during the second HS and subsequent recovery phase. The subcellular localization of YFP-HaFT-1 fusion protein was mainly detected in cytoplasm. HaFT-1 overexpression increased the germination rate of transgenic Arabidopsis seeds, and the survival rate of HaFT-1 overexpression seedlings was higher than that of wild-type (WT) Arabidopsis after priming-and-triggering and non-primed control treatments. Cell death staining showed that HaFT-1 overexpression lines exhibited significantly reduced cell death during HS compared to WT. Transcriptome analysis showed that genes associated with energy generation, protein metabolism, proline metabolism, autophagy, chlorophyll metabolism and reactive oxygen species (ROS) scavenging were important to the thermotolerance of HS-primed HaFT-1 transgenic plants. Growth physiology analysis indicated that priming-and-triggering treatment of Arabidopsis seedlings overexpressing HaFT-1 increased proline content and strengthened ROS scavenging activity. These results demonstrated that overexpression of HaFT-1 increased not only HS priming but also tolerance to the second HS of transgenic Arabidopsis, suggesting that HaFT-1 is a positive regulator in acquired thermotolerance.

Metabolomic Analysis of the Response of Haloxylon ammodendron and Haloxylon persicum to Drought.

Haloxylon ammodendron and Haloxylon persicum, as typical desert plants in arid areas, show strong drought tolerance and environmental adaptability and are therefore ideal model plants for studying the molecular mechanisms of drought tolerance. A metabolomic analysis of H. ammodendron and H. persicum in their natural environment is lacking, and their metabolic response to drought therefore remains unclear. To elucidate the response of H. ammodendron and H. persicum to drought at the metabolic level, a non-targeted metabolomics analysis was carried out herein. Under a dry environment, H. ammodendron exhibited 296 and 252 differentially expressed metabolites (DEMs) in the positive and negative ion modes, respectively, whereas 452 and 354 DEMs were identified in the positive and negative ion modes in H. persicum, respectively. The results indicated that H. ammodendron responds to drought by increasing the content of organic nitrogen compounds and lignans, neolignans, and related compounds, and reducing the content of alkaloids and derivatives. By contrast, H. persicum adapts to the dry environment by increasing the content of organic acids and their derivatives and reducing the content of lignans, neolignans, and related compounds. In addition, H. ammodendron and H. persicum improved their osmoregulation ability, reactive oxygen species detoxification ability, and cell membrane stability by regulating the key metabolic pathways and anabolism of associated metabolites. This is the first metabolomics report on the response of H. ammodendron and H. persicum to drought in their natural environment, providing a foundation for the further study of their regulatory mechanisms under drought stress.

Bioremediation of phenanthrene in saline-alkali soil by biochar- immobilized moderately halophilic bacteria combined with Suaeda salsa L.

Polycyclic aromatic hydrocarbon (PAH) contaminated saline-alkali soil is commonly salinized and hardened, which leads to low self-purification efficiency, making it difficult to reuse and remediate. In this study, pot experiments were conducted to investigate remediation of PAH contaminated saline-alkali soil using biochar-immobilized Martelella sp. AD-3, and Suaeda salsa L (S. salsa). Reduction in phenanthrene concentration, PAH degradation functional genes, and the microbial community in the soil were analyzed. The soil properties and plant growth parameters were also analyzed. After a 40-day remediation, the removal rate of phenanthrene by biochar-immobilized bacteria combined with S. salsa (MBP group) was 91.67 %. Additionally, soil pH and electrical conductivity (EC) reduced by 0.15 and 1.78 ds/m, respectively. The fresh weight and leaf pigment contents increased by 1.30 and 1.35 times, respectively, which effectively alleviated the growth pressure on S. salsa in PAH-contaminated saline-alkali soil. Furthermore, this remediation resulted in abundance of PAH degradation functional genes in the soil, with a value of 2.01 × 103 copies/g. The abundance of other PAH degraders such as Halomonas, Marinobacter, and Methylophaga in soil also increased. Furthermore, the highest abundance of Martelella genus was observed after the MBP treatment, indicating that strain AD-3 has a higher survival ability in the rhizosphere of S. salsa under the protection of biochar. This study provides a green, low-cost technique for remediation of PAH-contaminated saline-alkali soils.

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.

Biostimulation of Salicornia europaea L. crops with plant growth-promoting bacteria in laboratory and field conditions: effects on growth and metabolite profile.

AIM: The objective of the work was to assess the effect of biostimulation with selected plant growth-promoting bacteria on growth and metabolite profile of Salicornia europaea. METHODS AND RESULTS: Salicornia europaea seeds were inoculated with different combinations of plant growth-promoting bacteria Brevibacterium casei EB3, Pseudomonas oryzihabitans RL18, and Bacillus aryabhattai SP20. Plants germinated from inoculated seeds were grown either in laboratory conditions or in a saline crop field. Fresh and dry weight were determined at the end of the experiment, for biomass quantification. The microbiological quality of fresh shoots for human consumption as salad greens was assessed, and the persistence of the inoculated strains in the plant rhizosphere was confirmed by next-generation sequencing (Illumina) of the 16S rDNA gene. The primary metabolite profile of biostimulated plants was characterized by GC-TOF-MS.In laboratory conditions, inoculation with the two strains Br. casei EB3 and Ps. oryzihabitans RL18 caused the most significant increase in biomass production (fresh and dry weight), and caused a shift in the central metabolic pathways of inoculated plants toward amino acid biosynthesis. In the field experiment, no significant biostimulation effect was detected with any of the tested inoculants. Seed inoculation had no significant effect on the microbiological quality of the edible parts. The persistence of inoculants was confirmed in both experiments. CONCLUSIONS: Manipulation of the plant microbiome can trigger primary metabolic reconfiguration and modulate the plant metabolism while promoting plant growth.

Regulation of chromium translocation to shoot and physiological, metabolomic, and ionomic adjustments confer chromium stress tolerance in the halophyte Suaeda maritima.

Chromium (Cr) is a highly toxic element adversely affecting the environment, cultivable lands, and human populations. The present study investigated the effects of Cr (VI) (100-400 µM) on plant morphology and growth, photosynthetic pigments, organic osmolytes, ionomics, and metabolomic dynamics of the halophyte Suaeda maritima to decipher the Cr tolerance mechanisms. Cr exposure reduced the growth and biomass in S. maritima. The photosynthetic pigments content significantly declined at higher Cr concentrations (400 µM). However, at lower Cr concentrations (100-300 µM), the photosynthetic pigments remained unaffected or increased. The results suggest that a high concentration of Cr exposure might have adverse effects on PS II in S. maritima. The enhanced uptake of Na+ in S. maritima imposed to Cr stress indicates that Na+ might have a pivotal role in osmotic adjustment, thereby maintaining water status under Cr stress. The proline content was significantly upregulated in Cr-treated plants suggesting its role in maintaining osmotic balance and scavenging ROS. The metabolomic analysis of control and 400 µM Cr treated plants led to the identification of 62 metabolites. The fold chain analysis indicated the upregulation of several metabolites, including phytohormones (SA and GA3), polyphenols (cinnamic acid, sinapic acid, coumaric acid, vanillic acid, and syringic acid), and amino acids (alanine, leucine, proline, methionine, and cysteine) under Cr stress. The upregulation of these metabolites suggests the enhanced metal chelation and sequestration in vacuoles, reducing oxidative stress by scavenging ROS and promoting photosynthesis by maintaining the chloroplast membrane structure and photosynthetic pigments. Furthermore, in S. maritima, Cr tolerance index (Ti) was more than 60% in all the treatments, and Cr bio-concentration factor (BCF) and translocation factor (Tf) values were all greater than 1.0, which clearly indicates the Cr-hyperaccumulator characteristics of this halophyte.

Mitigation of salinity stress effects on kochia (Bassia scoparia L.) biomass productivity using biochar application.

Salinity continues to be a key factor limiting food security and agricultural sustainability in arid and semi-arid countries. Biochar has been promoted to reduce the risk of saline irrigation. In a controlled study, physiological and morphological growth factors of kochia plants that were irrigated with tap water (S1) and saline water (S2) were assessed to identify the ameliorative effects of biochar amendment to the soil at different levels (B1: 0%, B2: 2%, B3: 5%, and B4: 10%. According to our findings, salinity stress negatively affected morphological and physiological growth parameters of kochia plants by decreasing the fresh and dry weight (25% and 28%, respectively), plant height (30%), circumference (46%), total chlorophyll (51%), and relative water content (29%) when compared to the controls. Furthermore, electrolyte leakage increased considerably (19%) due to salt stress. Significant morphological and physiological growth enhancements were seen at all biochar levels in comparison to the control treatment, with the highest level increasing plant height by 55%, circumference by 76%, total chlorophyll concentrations by 121%, and relative water content by 28%. Furthermore, it resulted in a 36% reduction in the stressed plants' electrolyte leakage. The findings demonstrated biochar's benefits in reducing salinity's negative effects on kochia plants.

• This study provides new data about the specifying the impact of using biochar on salinity concentration and the growth parameters of kochia plants. This investigation demonstrated a significant results in terms of that the salinity stress relative to using biochar.• Biochar is crucial for stimulating and activating biochemical defensive mechanisms against salt stress; yet, research in this area is lacking.• Biochar has shown that it is crucial to stimulate biochemical defense mechanisms against salinity stress• It was found that using biochar as a soil supplement improved morphological, physiological, and biochemical characteristics of the kochia plant by increasing fresh and dried weight per plant, plant height, plant circumference, chlorophyll concentrations, and relative water content while lowering electrolyte leakage in stressed kochia plants. This research will aid in increasing kochia's early development and stand establishment in saline circumstances.

Chloride Channel Family in the Euhalophyte Suaeda altissima (L.) Pall: Cloning of Novel Members SaCLCa2 and SaCLCc2, General Characterization of the Family.

CLC family genes, comprising anion channels and anion/H+ antiporters, are widely represented in nearly all prokaryotes and eukaryotes. CLC proteins carry out a plethora of functions at the cellular level. Here the coding sequences of the SaCLCa2 and SaCLCc2 genes, homologous to Arabidopsis thaliana CLCa and CLCc, were cloned from the euhalophyte Suaeda altissima (L.) Pall. Both the genes cloned belong to the CLC family as supported by the presence of the key conserved motifs and glutamates inherent for CLC proteins. SaCLCa2 and SaCLCc2 were heterologously expressed in Saccharomyces cerevisiae GEF1 disrupted strain, Δgef1, where GEF1 encodes the only CLC family protein, the Cl− transporter Gef1p, in undisrupted strains of yeast. The Δgef1 strain is characterized by inability to grow on YPD yeast medium containing Mn2+ ions. Expression of SaCLCa2 in Δgef1 cells growing on this medium did not rescue the growth defect phenotype of the mutant. However, a partial growth restoration occurred when the Δgef1 strain was transformed by SaCLCa2(C544T), the gene encoding protein in which proline, specific for nitrate, was replaced with serine, specific for chloride, in the selectivity filter. Unlike SaCLCa2, expression of SaCLCc2 in Δgef1 resulted in a partial growth restoration under these conditions. Analysis of SaCLCa2 and SaCLCc2 expression in the euhalophyte Suaeda altissima (L.) Pall by quantitative real-time PCR (qRT-PCR) under different growth conditions demonstrated stimulation of SaCLCa2 expression by nitrate and stimulation of SaCLCc2 expression by chloride. The results of yeast complementation assay, the presence of both the "gating" and "proton" glutamates in aa sequences of both the proteins, as well results of the gene expression in euhalophyte Suaeda altissima (L.) Pall suggest that SaCLCa2 and SaCLCc2 function as anion/H+ antiporters with nitrate and chloride specificities, respectively. The general bioinformatic overview of seven CLC genes cloned from euhalophyte Suaeda altissima is given, together with results on their expression in roots and leaves under different levels of salinity.

Ion homeostasis in differently adapted populations of Suaeda vera Forssk. ex J.F. Gmel. for phytoremediation of hypersaline soils.

Salt-accumulator species are of great interest for the phytoremediation of salt-affected soils to reclaim soil salinization, a major constraints causing germination retardation and growth restriction of plants as well as habitat degradation. Higher biomass production at ECe 23-36 dS m-1 indicated that this species grows better in high to moderate salinity that was linked to osmotic adjustment through higher ion accumulation (Na+, Cl‒, and Ca2+) and organic osmolytes (free amino acids and proline). Plants from highly and moderately saline habitats exhibited broader metaxylem vessels, which was associated with eased conduction of solutes leading to better growth. Leaf anatomical characteristics generally increased with increasing salinity except at the highest ECe 55 dS m-1. The increased leaf lamina thickness contributed to succulence because of increased storage parenchymatous spongy tissues (that can store high amounts of water), water contents and it is a reflection of maintaining ion homeostasis and colonizing hyper-saline soil. Reduced stomatal density and area under high salinity are critical to cope with environmental hazards. Under high salinity, compartmentalization of excessive Na+ and Cl- ions and accumulation of compatible osmolytes are directly related to high degree of salinity tolerance, and hence are useful for phyto-amelioration of salinity-impacted lands.

Salinity tolerance mechanism in Suaeda vera have been explored extensively, but these mechanisms have not been addressed in the context of soil-plant interaction and functioning connection between structural and functional features with respect to phytoremediation. This work aims to provide mechanistic insight into how different morphological and physio-anatomical mechanisms in differently adapted population of Suaeda vera operate in coordinated manner to maintain ionic homeostasis under high salinity that are useful for phytoremediation.