Physiological and metabolic adjustments in the xero-halophyte Haloxylon salicornicum conferring drought tolerance.

Drought is one of the most catastrophic abiotic stresses that affects global food production severely. The present work investigates the metabolic and physiological adaptation mechanisms in the xero-halophyte Haloxylon salicornicum to counter the effects of drought. This xero-halophyte can withstand a prolonged drought period of 14 days and recovered within 7 days of irrigation with minimal effects of drought on growth and physiological parameters. Photosynthetic parameters such as PN , gs , and E decreased significantly, whereas WUE increased under drought condition. Drought induces a significant decline in the Fv/Fm ratio. However, the value of Fv/Fm ratio successfully recovered within 7 days of the recovery period. Differential regulations of various antioxidative enzymes increase the drought tolerance potential of H. salicornicum. The metabolomic analysis of H. salicornicum shoot identified 63 metabolites: 43 significantly increased and 20 significantly decreased under drought conditions. These metabolites mainly include amino acids, organic acids, amines, sugar alcohols, sugars, fatty acids, alkaloids, and phytohormones. The metabolites that have a significant contribution towards drought tolerance include citric acid, malic acid, tartaric acid, d-erythrose, glyceric acid, sucrose, pentanoic acid, d-mannitol, ABA, and palmitic acid. KEGG pathway enrichment analysis showed that the vital drought-responsive metabolic pathways mainly include galactose metabolism, aminoacyl-tRNA biosynthesis, glyoxylate and dicarboxylate metabolism, citrate cycle (TCA cycle), alanine, aspartate, and glutamate metabolism. This study offers comprehensive information on physiological, antioxidative and metabolic adaptations and overall drought tolerance mechanisms in H. salicornicum. The information gained from this study will provide guidance to plant breeders and molecular biologists to develop drought-tolerant crop varieties.

Unraveling salt responsive metabolites and metabolic pathways using non-targeted metabolomics approach and elucidation of salt tolerance mechanisms in the xero-halophyte Haloxylon salicornicum.

Haloxylon salicornicum is a xero-halophyte growing in saline and arid regions of the world. Metabolite profiling was carried out in shoot of both control and salinity treated (400 mM NaCl) samples by GC-QTOF-MS and HPLC-DAD analysis to decipher the salinity tolerance mechanism in this xero-halophyte. The present study investigates the alteration in metabolite profile of H. salicornicum that support the salinity tolerance of the plant. The metabolomic analysis of H. salicornicum shoot identified 56 metabolites, of which 47 metabolites were significantly changed in response to salinity. These metabolites were mainly included in the category of amino acids, organic acids, amines, sugar alcohols, sugars, fatty acids, alkaloids, and phytohormones. In response to salinity, most of the amino acids were down-regulated except alanine, phenylalanine, lysine, and tyramine, which were up-regulated in H. salicornicum. In contrast to amino acids, most sugars and organic acids were up-regulated in response to salinity. Correlation and pathway enrichment analysis identified important biological pathways playing significant roles in conferring salt tolerance of H. salicornicum. These biological pathways include amino sugar and nucleotide sugar metabolism, citrate cycle (TCA cycle), starch and sucrose metabolism, phenylalanine metabolism, cysteine, methionine, glycine, serine, and threonine metabolism, etc. The data presented here suggest that the modulations of various metabolic pathways facilitate H. salicornicum to survive and grow optimally even under high salinity condition. This study offers comprehensive information on metabolic adaptations and overall salt tolerance mechanisms in H. salicornicum. The information gained through this study will provide guidance to plant breeders and molecular biologists to develop salinity tolerant crop varieties.

Comprehensive proteomic analysis revealing multifaceted regulatory network of the xero-halophyte Haloxylon salicornicum involved in salt tolerance.

Haloxylon salicornicum is a xero-halophyte which grow predominantly in dry saline areas. However, the proteomic approach for revealing the regulatory network involved in salt adaptation of this important xerohalophyte has not been studied so far. In the present investigation, the label-free quantitative proteomic analysis was carried out in shoot of H. salicornicum to get an insight into the functional network of proteins involved in salt tolerance. Comparative proteomic analysis in control and salt treated plants of H. salicornicum by nano-ESI-LC-MS and MS/MS, and data base searching led to the identification of 723 proteins. Pathway enrichment analysis by KEGG uncovered various biological pathways to which salinity induced differentially regulated proteins are involved. In H. salicornicum, out of 723 identified proteins, 187 proteins were differentially regulated in response to salinity. In addition to significant up-regulation of stress responsive proteins, other proteins involved in carbohydrate metabolism, TCA cycle, protein synthesis, antioxidative defense systems, energy transfer, ion transport, nucleotide binding, and proteosomal proteins also significantly up-regulated under salinity in H. salicornicum. The major photosynthetic proteins up-regulated were RuBisCo, D1 protein, photosystem II-CP47, and cytochrome b599. TCA cycle component proteins such as citrate synthase, succinate dehydrogenase, and malate dehydrogenase upregulated indicating their significant roles in providing vital energy for salinity tolerance. Salinity induced higher expressions of ion transporters in H. salicornicum suggest efficient compartmentalization of toxic sodium ions. In addition, up-regulation of antioxidative defense system can be correlated with effective scavenging of salinity induced ROS, hence imparting salt tolerance. In H. salicornicum, protein synthesis was boosted under salinity as confirmed from the salinity-induced up-regulation of the ribosome associated proteins. Salinity induced significantly changed proteins of the ribosomal pathway include ribosomal protein components such as elongation factor-Tu (EF-Tu), initiation factor 1 and 2 (IF1, 2), Rpo cluster C and B, etc. Functional integrity of protein synthesis machinery in H. salicornicum is maintained under high salinity by higher abundance of ribosomal subunit proteins in NaCl-treated plants. We assume that consistent energy supply by the up-regulation of TCA cycle along with uninterrupted protein synthesis and maintenance of structural integrity of the photosynthetic machinery are the primary mechanism of salinity tolerance of H. salicornicum. In the present study, we comprehensively elucidated possible mechanisms associated with systematic salt tolerance of H. salicornicum employing proteomic approach. The information from this study will contribute to the genetic improvement of crop plants that can be grown in saline marginal lands.

Mineral nutrient homeostasis, photosynthetic performance, and modulations of antioxidative defense components in two contrasting genotypes of Arachis hypogaea L. (peanut) for mitigation of nitrogen and/or phosphorus starvation.

Arachis hypogaea L. (peanut) is a major oil yielding crop and its productivity is largely affected by the availability of nitrogen and phosphorus. The present study aims to elucidate the differential physiological and biochemical mechanisms involved in two contrasting genotypes of peanut for mitigation of N and/or P deficiency. The plants of two contrasting genotypes of peanut (GG7 and TG26) were subjected to N and/or P deficiency under hydroponic culture condition. After 15 d of N and/or P deficiency, various growth parameters, mineral nutrient status, nutrient use efficiency, photosynthesis, transpiration, water use efficiency, chlorophyll fluorescence, ROS level, and changes in enzymatic and non-enzymatic antioxidative components were measured in control and nutrient deficient plants. Our results showed that GG7 is fast-growing genotype than TG26 under control condition, whereas under N and/or P deficiency growth performance of GG7 was significantly declined as compared to TG26. The levels of photosynthetic pigments, net photosynthesis activity (PN), and stomatal conductance (gs) declined in N and/or P deficient plants of both the genotypes. However, quantum efficiency of photosystem II (Fv/Fm) did not change significantly under N and/or P starvation in both the genotypes. In the present investigation, most of the antioxidative enzymes either remained in steady state or downregulated in both the genotypes of peanut under N and/or P deficiency condition. N and/or P deficiency did not influence the levels of ROS and oxidative stress indicators such as O2·-, H2O2, and MDA in both the genotypes. In the present investigation, the decline in growth in both the genotypes under N and/or P deficiency might be due to the reduced photosynthetic performance. Our results suggest that TG26 is more resistant to N and P deficiency than GG7 genotype. Higher NUE value of GG7 as compared to TG26 suggests that GG7 can utilize N more efficiently to promote biomass production than TG26 under sufficient nutrient condition. On the other hand, mineral resource allocation to leaf and higher PUE are key adaptive features of the TG26 genotype under N, and P deficiency conditions. The differential regulations of various enzymatic and non-enzymatic antioxidative components in peanut genotypes maintain the cellular redox homeostasis under mineral deficiency conditions and prevent the peanut plants from oxidative stress, thereby maintaining PSII efficiency. The information from the present study can be useful for the improvement of traits in peanut that can maintain the productivity under N and P deficient environment with minimum input of fertilizers.

Metabolomic study reveals key metabolic adjustments in the xerohalophyte Salvadora persica L. during adaptation to water deficit and subsequent recovery conditions.

Water deficit severely limits productivity of plants, and pose a major threat to modern agriculture system. Therefore, understanding drought adaptive mechanisms in drought-tolerant plants is imperative to formulate strategies for development of desiccation tolerance in crop plants. In present investigation, metabolic profiling employing GC-QTOF-MS/MS and HPLC-DAD was carried out to evaluate metabolic adjustments under drought stress in the xero-halophyte Salvadora persica. The metabolite profiling identified a total of 68 metabolites in S. persica leaf, including organic acids, amino acids, sugars, sugar alcohols, hormones, and polyphenols. The results showed that higher cellular osmolality under drought stress was accompanied by accumulations of several osmoprotectants like sugars and polyols (sucrose, glucose, mannose, galactose, erythrose, sorbose, glycerol, and myoinositol), organic acids (galactaric acid, tartaric acid, malic acid, oxalic acid, and citric acid), and amino acids (alanine, phenylalanine, tyrosine). Upregulation of ABA and JA support to achieve early drought tolerance in S. persica. Moreover, accumulation of coumarin, gallic acid, and chlorogenic acid provide antioxidative defense to S. persica. KEGG pathway enrichment analysis showed that altered metabolites were associated with starch and sucrose metabolism, galactose metabolism, inositol phosphate metabolism, and phenylalanine metabolism. While during recovery, metabolites associated with lysine biosynthesis and alanine, aspartate and glutamate metabolism were significantly altered. The results of the present study imply that coordinated regulations between various metabolites, metabolic processes, and pathways empower the xerohalophyte S. persica to adapt under drought environment. The knowledge from this study will enable the development of drought tolerance in crops using genetic engineering and breeding approaches.

Phytochemical profiling, polyphenol composition, and antioxidant activity of the leaf extract from the medicinal halophyte Thespesia populnea reveal a potential source of bioactive compounds and nutraceuticals.

The present study evaluated the phytochemical constituents, nutritional attributes, and the antioxidant capacity of the medicinal halophyte Thespesia populnea. The metabolite profiling by GC-QTOF-MS analysis identified 37 metabolites among which sucrose, malic acid, and turanose were the most abundant. A total of 18 polyphenols and 17 amino acids were identified by the HPLC-DAD analysis. The most abundant polyphenols in T. populnea were gallic acid, catechin, and myricetin. Other polyphenols like protocatechuic acid, epigallocatechin gallate, rosmarinic acid, ellagic acid, rutin, and naringenine were also detected in ample amounts. The leaf extract demonstrated higher antioxidant as well as lipid peroxidation inhibition activities. A correlation analysis revealed a positive correlation between the antioxidant capacity and the phenolic compounds viz. gallic acid, catechin, myricetin, quercetin, apigenin, cinnamic acid, and coumarin which indicates that these phenolic compounds are the main contributors of the antioxidant potential of T. populnea. The results of this study establish T. populnea as a potential source of nonconventional functional food. PRACTICAL APPLICATIONS: The data presented here indicate that T. populnea can be considered as a nonconventional functional food and potential source of energy, antioxidants, minerals, essential amino acids, and bioactive compounds in herbal formulations, food supplements, or nutraceuticals. The metabolites identified from this halophyte have pharmacological and nutraceutical potentials, suggesting T. populnea as an ideal candidate for application in the food and phytopharmaceutical industries to produce health-promoting products, functional foods, and herbal medicines.

Regulation of ROS through proficient modulations of antioxidative defense system maintains the structural and functional integrity of photosynthetic apparatus and confers drought tolerance in the facultative halophyte Salvadora persica L.

The facultative halophyte Salvadora persica L. grow in arid, semiarid and saline areas. In present study, drought induced alterations in growth, ion homeostasis, photosynthesis, chlorophyll fluorescence, ROS regulation and antioxidative defense components were analyzed in S. persica with an aim to elucidate the drought tolerance mechanisms. In response to drought, significant reductions in growth, photosynthesis, and photosynthetic pigments were observed in S. persica. However, leaf relative water content (RWC %) did not change significantly. In S. persica seedlings, the growth, photosynthetic pigment contents and photosynthesis were resumed to control level within 7 d, when the drought treated plants were re-irrigated. However, quantum yield of PSII (ΦPSII), rate of electron transport (ETR), maximum efficiency of PSII (Fv/Fm), and photochemical quenching (qP) remained unaffected under water deficit stress. The results suggest that both non-stomatal as well as stomatal limitations can account for photosynthetic reduction. The ionomics studies revealed no significant alterations in levels of Na+, K+, Ca2+, B, Cu2+, Fe2+, Mo, and Zn2+ in leaf tissue during drought. However, there was increase in levels of Na+, K+, Ca2+ and Mg2+ in root tissue in response to drought. The activity of different enzymatic antioxidants like SOD, APX, and GR remained unaffected during drought, whereas POX activity increased and CAT activity declined under drought stress in comparison to control. This result proposes that vital ROS scavenging enzymes like SOD, APX and GR are at threshold levels to maintain the appropriate concentration of ROS. In S. persica, the ratio of AsA/DHA and GSH/GSSG (which are the indicators of redox potential of cell) remained steady or increased under drought which indicates that cellular redox level is maintained in this halophyte. Although ROS levels (H2O2 and O2•-) increased significantly under drought stress, electrolyte leakage and lipid peroxidation level remained unchanged in response to water deficit condition which indicates that minimal increase in ROS level under drought stress act in signaling for activation of ROS scavenging enzymes. Our results propose that decline in growth and photosynthesis is a vital energy conservation strategy of S. persica under drought condition. The rapid recovery of growth, photosynthesis and water relations in S. persica following drought seems to be a critical mechanism permitting this plant to withstand and survive under drought environment. In addition, our results implicate that efficient regulations of antioxidative enzymes in leaf tissue contribute in regulating the ROS level and cellular redox status, thereby protecting the plant from drought induced oxidative damage in S. persica. Consequently ion homeostasis, plant water status, and integrity of photosynthetic apparatus is maintained in S. persica subjected to drought. The results of present study propose that S. persica is a drought tolerant halophyte and it can be a potential candidate for restoration of degraded saline lands of coastal ecosystem.

Antioxidant Activities, Metabolic Profiling, Proximate Analysis, Mineral Nutrient Composition of Salvadora persica Fruit Unravel a Potential Functional Food and a Natural Source of Pharmaceuticals.

Salvadora persica is a medicinally important plant mainly used in oral hygiene. However, little attention has been given towards the nutritional prominence of this plant. This study encloses the proximate and mineral nutrient contents, amino acid composition, metabolite profiling and antioxidant potential of S. persica fruit. The ripen fruit contained substantial amount of sugars, mineral nutrients, carotenoids, polyphenols and flavonoids. The metabolic profiling of the fruit extract by GC-MS revealed a total of 22 metabolites comprising of sugars, sugar alcohols, organic acids, organic base, and aromatic silica compound. The identified metabolites have been previously reported to have potential antioxidant, antimicrobial, anti-hyperglycemic, and antitumor properties. The GC-MS analysis indicated high glucose and glucopyranose (247.62 and 42.90 mg g-1 FW respectively) contents in fruit of S. persica. The fruit extract demonstrated a significantly higher antioxidant and ROS scavenging properties along with high contents of mineral nutrients and essential amino acids. HPLC analysis revealed presence of essential and non-essential amino acid required for healthy body metabolism. The cysteine was found to be in highest amount (733.69 mg 100 g-1 DW) among all amino acids quantified. Specifically, compared to similar medicinal plants, previously reported as a source of non-conventional food and with some of the commercially important fruits, S. persica fruit appears to be a potential source of essential mineral nutrients, amino acids, vitamins (ascorbic acid and carotenoid) and pharmaceutically important metabolites contributing towards fulfilling the recommended daily requirement of these for a healthy human being. This is the first report establishing importance of S. persica fruit as nutraceuticals. The data presented here proposed that fruit of S. persica may be used as functional food or reinvigorating ingredient for processed food to reduce deficiency of nutrients among the vulnerable population group. The phytochemicals identified from S. persica fruit may be used as natural source for pharmaceutical preparations.

Coordinated Changes in Antioxidative Enzymes Protect the Photosynthetic Machinery from Salinity Induced Oxidative Damage and Confer Salt Tolerance in an Extreme Halophyte Salvadora persica L.

Salinity-induced modulations in growth, photosynthetic pigments, relative water content (RWC), lipid peroxidation, photosynthesis, photosystem II efficiency, and changes in activity of various antioxidative enzymes were studied in the halophyte Salvadora persica treated with various levels of salinity (0, 250, 500, 750, and 1000 mM NaCl) to obtain an insight into the salt tolerance ability of this halophyte. Both fresh and dry biomass as well as leaf area (LA) declined at all levels of salinity whereas salinity caused an increase in leaf succulence. A gradual increase was observed in the Na(+) content of leaf with increasing salt concentration up to 750 mM NaCl, but at higher salt concentration (1000 mM NaCl), the Na(+) content surprisingly dropped down to the level of 250 mM NaCl. The chlorophyll and carotenoid contents of the leaf remained unaffected by salinity. The photosynthetic rate (PN), stomatal conductance (gs), the transpiration rate (E), quantum yield of PSII (ΦPSII), photochemical quenching (qP), and electron transport rate remained unchanged at low salinity (250 to 500 mM NaCl) whereas, significant reduction in these parameters were observed at high salinity (750 to 1000 mM NaCl). The RWC% and water use efficiency (WUE) of leaf remained unaffected by salinity. The salinity had no effect on maximum quantum efficiency of PS II (Fv/Fm) which indicates that PS II is not perturbed by salinity-induced oxidative damage. Analysis of the isoforms of antioxidative enzymes revealed that the leaves of S. persica have two isoforms each of Mn-SOD and Fe-SOD and one isoform of Cu-Zn SOD, three isoforms of POX, two isoforms of APX and one isoform of CAT. There was differential responses in activity and expression of different isoforms of various antioxidative enzymes. The malondialdehyde (MDA) content (a product of lipid peroxidation) of leaf remained unchanged in S. persica treated with various levels of salinity. Our results suggest that the absence of pigment degradation, the reduction of water loss, and the maintenance of WUE and protection of PSII from salinity-induced oxidative damage by the coordinated changes in antioxidative enzymes are important factors responsible for salt tolerance of S. persica.