Growth, Osmolyte Concentration and Antioxidant Enzymes in the Leaves of Sesuvium portulacastrum L. Under Salinity Stress

In this study, growth and osmolyte concentration in the leaves of halophyte, Sesuvium portulacastrum, were studied with respect to salinity. Therefore, the changes in shoot growth, leaf tissue water content, osmolyte concentration (proline content, glycine betaine) and antioxidant enzymes [polyphenol oxidase (PPO), superoxide dismutase (SOD) and catalase (CAT)] were investigated. The 30-day old S. portulacastrum plants were subjected to 100, 200, 300, 400, 500 and 600 mM NaCl for 28 days. The plant growth was steadily increased up to 500 mM NaCl stress at 28 days. TWC was higher in 300 mM NaCl treated leaves than that of 600 mM NaCl. Salinity stress induced the accumulation of osmolyte concentration when compared to control during the study period. The antioxidant enzymes PPO, CAT and SOD were increased under salinity.     

Exogenous Sodium Nitroprusside Mitigates Salt Stress in Lentil (Lens culinaris Medik.) by Affecting the Growth,Yield, and Biochemical Properties

Soil salinity disrupts the physiological and biochemical processes of crop plants and ultimately leads to compromising future food security. Sodium nitroprusside (SNP), a contributor to nitric oxide (NO), holds the potential to alleviate abiotic stress effects and boost tolerance in plants, whereas less information is available on its role in salt-stressed lentils. We examined the effect of exogenously applied SNP on salt-stressed lentil plants by monitoring plant growth and yield-related attributes, biochemistry of enzymes (superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD)) amassing of leaf malondialdehyde (MDA) and hydrogen peroxide (H₂O₂). Salinity stress was induced by NaCl application at concentrations of 50 mM (moderate salinity) and 100 mM (severe salinity), while it was alleviated by SNP application at concentrations of 50 µM and 100 µM. Salinity stress severely inhibited the length of roots and shoots, the relative water content, and the chlorophyll content of the leaves, the number of branches, pods, seeds, seed yield, and biomass per plant. In addition, MDA, H₂O₂ as well as SOD, CAT, and POD activities were increased with increasing salinity levels. Plants supplemented with SNP (100 µM) showed a significant improvement in the growth- and yield-contributing parameters, especially in plants grown under moderate salinity (50 mM NaCl). Essentially, the application of 100 µM SNP remained effective to rescue lentil plants under moderate salinity by regulating plant growth and biochemical pathways. Thus, the exogenous application of SNP could be developed as a useful strategy for improving the performance of lentil plants in salinity-prone environments.     

RAPD Markers Associated with Salt Tolerance in Soybean Genotypes Under Salt Stress

In order to investigate the influence of genetic background on salt tolerance in soybean (Glycine max), ten soybean genotypes (Pusa-20, Pusa-40, Pusa-37, Pusa-16, Pusa-24, Pusa-22, BRAGG, PK-416, PK-1042, and DS-9712) released in India, were selected and grown hydroponically. The 10-day-old seedlings were subjected to 0, 25, 50, 75, 100, 125, and 150 mM NaCl for 15 days. Plant growth, leaf osmotic adjustment, and random amplified polymorphic DNA (RAPD) analysis were studied. In comparison to control plants, the plant growth in all genotypes was decreased by salt stress, respectively. Salt stress decreased leaf osmotic potential in all genotypes; however, the maximum reduction was observed in genotype Pusa-24 followed by PK-416 and Pusa-20, while minimum reduction was shown by genotype Pusa-37, followed by BRAGG and PK-1042. Pusa-16, Pusa-22, Pusa-40, and DS-9712 were able to tolerate NaCl treatment up to the level of 75 Mm. The difference in osmotic adjustment between all the genotypes was correlated with the concentrations of ion examined such as Na⁺ and the leaf proline concentration. These results suggest that the genotypic variation for salt tolerance can be partially accounted by plant physiological measures. Twenty RAPD primers revealed high polymorphism and genetic variation among ten soybean genotypes studied. The closer varieties in the cluster behaved similarly in their response to salinity tolerance. Intra-clustering within the two clusters precisely grouped the ten genotypes in sub-cluster as expected from their physiological findings. Our study shows that RAPD technique is a sensitive, precise, and efficient tool for genomic analysis in soybean genotypes.     

Lipoic Acid Combined with Melatonin Mitigates Oxidative Stress and Promotes Root Formation and Growth in Salt-Stressed Canola Seedlings (Brassica napus L.)

Lipoic acid (LA) and melatonin (MT) are pleiotropic molecules participating in plant stress resistance by modulating cellular biochemical changes, ion homeostasis, and antioxidant enzyme activities. However, the combined role of these two molecules in counteracting the detrimental impacts of salinity stress is still unknown. In the present study, we determined the effects of exogenous LA (0.5 µM), MT (1 µM) and their combination (LA + MT) on growth performance and biomass accumulation, photosynthetic pigments, enzymatic and non-enzymatic antioxidant activities, and ions homeostatic in canola (Brassica napus L.) seedlings under salinity stress (0, 100 mM) for 40 days. The results indicate that exogenous application of LA + MT improved the phenotypic growth (by 25 to 45%), root thickness (by 68%), number of later lateral roots (by 52%), root viability (by 44%), and root length (by 50%) under salinity stress. Moreover, total soluble protein, chlorophyll pigments, the concentration of superoxide dismutase (SOD), catalase peroxidase (CAT), and ascorbic peroxidase (ASA) increased with the presence of salt concentration into the growth media and then decreased with the addition of LA + MT to saline solution. Leaf protein contents and the degradation of photosynthetic pigments were lower when LA + MT treatments were added into NaCl media. The proline and phenol contents decreased in the exogenous application of LA + MT treatments more than individual LA or MT treatments under the salinity stress. The incorporation of LA or MT or a combination of LA + MT to saline solution decreased salinity-induced malondialdehyde and electrolyte leakage. In conclusion, the alteration of metabolic pathways, redox modulation, and ions homeostasis in plant tissues by the combined LA and MT application are helpful towards the adaptation of Brassica napus L. seedlings in a saline environment. The results of this study provide, for the first time, conclusive evidence about the protective role of exogenous LA + MT in canola seedlings under salinity stress.     

Effects of Saline-Alkaline Stress on Seed Germination and Seedling Growth of Sorghum bicolor (L.) Moench

In order to study the adaptation ability of sweet sorghum (Sorghum bicolor L. Moench) in the Yellow River Delta, the sweet sorghum variety Mart was used in this study to determine the roles of different saline-alkaline ratio stress treatment during seed germination to seedling stage. The results showed that Na⁺ concentration had a significant impact on the seed germination, seedling growth, and plant survival of sweet sorghum. Increasing Na⁺ concentration led to a decline in germination rate, final germination percentage, survival percentage, plant height, and dry weight per plant, a prolonged mean time of germination, as well as loss of improvement effect of low-Na+ concentration. The interaction effect of Na⁺ concentration and pH on the mean time of germination and germination rate was not significant (p?+ concentration (100 mM), high pH reduced the mean time of germination and increased the germination rate, without decline in final germination percentage and survival percentage. Therefore, at least in the duration of seed germination to the harvest period in the research, the sweet sorghum was resistant to the pH stress (≥9.04) when the Na⁺ concentration was below 100 mM. When suffered from the saline-alkaline stress, the seedling of sweet sorghum was characterized by ecological adaptive features, such as decreased stem ratio and chlorophyll b content in leaves and increased root ratio and chlorophyll a content, in order to maintain the uptakes of water and nutrient, and carbon assimilation. When the stress intensified, the lipid oxidation products, e.g., malondialdehyde (MDA), increased in sweet sorghum seedlings. However, the increasing of soluble protein content and antioxidant enzyme activity (superoxide dismutase (SOD), guaiacol peroxidase (POD), and gatalase (CAT)) was only founded in neutral low-Na⁺ concentration treatment (A₁), which indicated that high-salt concentration and pH all elicited harmful effects and limited the self-healing ability of sweet sorghum seedlings. In all, in order to grow sweet sorghum in the saline-alkaline soils of the Yellow River Delta, the salt concentration and pH value of the soil must be taken into consideration, and seeding density should be increased and supported by appropriate irrigation measures to reduce saline-alkaline stress so as to ensure the survival and growth of sweet sorghum seedlings.     

Protection of Vigna unguiculata (L.) Walp. plants from salt stress by paclobutrazol

A pot culture experiment was conducted to estimate the stress ameliorating ability of paclobutrazol, a triazole fungicide in Vigna unguiculata (L.) Walp. plants. Treatments were given as 80 mM NaCl, 80 mM NaCl + 15 mg l⁻¹ paclobutrazol and 15 mg l⁻¹ paclobutrazol alone. The samples were collected on 60 and 80 days after sowing (DAS). NaCl stress inhibited the root and stem length, total leaf area, fresh weight (FW), dry weight (DW) and activities of antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX). Plants treated with NaCl with paclobutrazol increased these parameters to a larger extent when compared to NaCl stressed plants. The results showed that the paclobutrazol significantly ameliorated the adverse effects of NaCl stress in V. unguiculata plants.     

Physiological and Biochemical Parameters of Common Duckweed Lemna minor after the Exposure to Tetracycline and the Recovery from This Stress

In this study, the ability of Lemna minor L. to recover to normal growth, after being degraded in a tetracycline-containing medium, was extensively investigated. The plants were exposed to tetracycline (TC) at concentrations of 1, 2.5, and 10 mM. Subsequently, their physiological status was analysed against the following criteria: rate of plant growth; free radical accumulation; antioxidant enzyme activity; chlorophyll content; HSP70 protein content; cell membrane permeability, and mitochondrial activity. The study showed that duckweed can considerably recover from the damage caused by antibiotics, within a week of cessation of stress. Of the plant properties analysed, mitochondrial activity was the most sensitive to antibiotic-induced disturbances. After transferring the plants to a tetracycline-free medium, all plant parameters improved significantly, except for the mitochondrial activity in the plants grown on the medium containing the highest dose of tetracycline. In the plants treated with this antibiotic at the concentration of 10 mM, the proportion of dead mitochondria increased and was as high as 93% after one week from the beginning of the recovery phase, even after the transfer to the tetracycline-free medium.     

An Overview of the Genetics of Plant Response to Salt Stress: Present Status and the Way Forward

Salinity is one of the major threats faced by the modern agriculture today. It causes multidimensional effects on plants. These effects depend upon the plant growth stage, intensity, and duration of the stress. All these lead to stunted growth and reduced yield, ultimately inducing economic loss to the farming community in particular and to the country in general. The soil conditions of agricultural land are deteriorating at an alarming rate. Plants assess the stress conditions, transmit the specific stress signals, and then initiate the response against that stress. A more complete understanding of plant response mechanisms and their practical incorporation in crop improvement is an essential step towards achieving the goal of sustainable agricultural development. Literature survey shows that investigations of plant stresses response mechanism are the focus area of research for plant scientists. Although these efforts lead to reveal different plant response mechanisms against salt stress, yet many questions still need to be answered to get a clear picture of plant strategy to cope with salt stress. Moreover, these studies have indicated the presence of a complicated network of different integrated pathways. In order to work in a progressive way, a review of current knowledge is critical. Therefore, this review aims to provide an overview of our understanding of plant response to salt stress and to indicate some important yet unexplored dynamics to improve our knowledge that could ultimately lead towards crop improvement.     

Genotypic Variability Among Soybean Genotypes Under NaCl Stress and Proteome Analysis of Salt-Tolerant Genotype

The present investigation was conducted to evaluate salt tolerance in ten genotypes of soybean (Glycine max L.). Twelve-day-old seedlings, grown hydroponically, were treated with 0, 25, 50, 75, 100, 125 and 150?mM NaCl for 10?days. Growth, lipid peroxidation and antioxidant enzyme activities were evaluated. Growth, measured in terms of length, fresh weight and dry weight of plants, was drastically reduced in Pusa-24 while there was little effect of NaCl treatment on Pusa-37 genotype of soybean. High level of lipid peroxidation was observed in Pusa-24 as indicated by increased level of malondialdehyde. Activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase were maximum in Pusa-37 where 9-, 1-, 5- and 6-fold increase over control were observed, respectively. The results suggested that Pusa-24 and Pusa-37 are salt-sensitive and salt-tolerant genotype of soybean, respectively, and antioxidant defence system is involved in conferring the sensitiveness and tolerance in these genotypes. Salt-tolerant genotype Pusa-37, was further analysed by 2-dimensional gel electrophoresis to analyse the differential expression of proteins at high salt stress. In the present study, 173 protein spots were identified. Of these, 40 proteins were responsive to salinity in that they were either up- or downregulated. This study could help us in identifying the possible regulatory switches (gene/s) controlling novel proteins of the salt-tolerant genotype of the crop plants and their possible role in defence mechanism.     

J-Aggregates Formed by NaCl Treatment of Aza-Coating Heptamethine Cyanines and Their Application to Monitoring Salt Stress of Plants and Promoting Photothermal Therapy of Tumors

The cationic nature of heptamethine cyanines gives them the capacity to form aggregates with salts by electrostatic interactions. In this work, NaCl promoted J-aggregate formation of aza-coating heptamethine cyanines is explored. NaCl can induce the N-benzyloxycarbonyl Cy-CO₂Bz to assemble into a J-aggregate having an absorption at 890 nm. Its excellent fluorescence response to NaCl implies that it has great potential for use as a probe for tracing salt stress in plants. Moreover, NaCl also promotes formation of J-aggregates from the N-ethyloxycarbonyl Cy-CO₂Et . The aggregate shows an intense absorption at 910 nm compared to the monomer which absorbs at 766 nm. Its J-aggregated form can serve as a photothermal agent. And the photothermal conversion efficiency is increased from 29.37 % to 57.59 %. This effort leads to the development of two applications of new cyanine J-aggregates including one for tracing salt stress of plants and the other for promoting photothermal therapy of tumors.     

Nutritional and Functional Evaluation of Inula crithmoides and Mesembryanthemum nodiflorum Grown in Different Salinities for Human Consumption

The nutritional composition and productivity of halophytes is strongly related to the biotic/abiotic stress to which these extremophile salt tolerant plants are subjected during their cultivation cycle. In this study, two commercial halophyte species (Inula crithmoides and Mesembryanthemum nodiflorum) were cultivated at six levels of salinity using a soilless cultivation system. In this way, it was possible to understand the response mechanisms of these halophytes to salt stress. The relative productivity decreased from the salinities of 110 and 200 mmol L⁻¹ upwards for I. crithmoides and M. nodiflorum, respectively. Nonetheless, the nutritional profile for human consumption remained balanced. In general, I. crithmoides vitamin (B1 and B6) contents were significantly higher than those of M. nodiflorum. For both species, β-carotene and lutein were induced by salinity, possibly as a response to oxidative stress. Phenolic compounds were more abundant in plants cultivated at lower salinities, while the antioxidant activity increased as a response to salt stress. Sensory characteristics were evaluated by a panel of culinary chefs showing a preference for plants grown at the salt concentration of 350 mmol L⁻¹. In summary, salinity stress was effective in boosting important nutritional components in these species, and the soilless system promotes the sustainable and safe production of halophyte plants for human consumption.     

Pseudomonas stutzeri Strain Possessing a Self-Transmissible TOL-Like Plasmid Degrades Phenol and Promotes Maize Growth in Contaminated Environments

Phenol is volatile organic pollutant that plants can little degrade. For complete degradation of volatile pollutants, we introduced Pseudomonas stutzeri strain P7 to phenol-contaminated soils. The strain effectively degraded phenol and even promoted plant growth. A TOL-like plasmid was detected in the strain and found to be responsible for phenol degradation and self-transmissible. In addition, phenol degradation by strain P7 was more rapid in the contaminated soils with than without plants over the full course of the experiment; especially by 5 days, the phenol concentration was reduced by about 30 % in soil without plants and reduced by about 50–65 % in soil with plants. This situation also occurred when inoculated with different transconjugants. Furthermore, transfer frequencies of TOL-like plasmid were significantly higher in soil with than without plants. Populations of rifampin-resistant P7 strain remained relatively constant for 20 days, while the number of rhizosphere bacteria that contained the degradative plasmids gradually increased at the later stages, suggesting that plants might stimulate plasmid transfer from strain P7 to indigenous bacteria, one possible reason for plant enhancing microbial degradation. This is attractive for implementation of combinations of phytoremediation and bioaugmentation in degradation of volatile pollutants that plants can little degrade.     

A hydroponic rice seedling culture model system for investigating proteome of salt stress in rice leaf

By using an in vivo hydroponic rice seedling culture system, we investigated the physiological and biochemical responses of a model rice japonica cultivar Nipponbare to salt stress using proteomics and classical biochemical methods. Yoshida's nutrient solution (YS) was used to grow rice seedlings. YS-grown 18-day-old seedlings manifested highly stable and reproducible symptoms, prominently the wilting and browning of the 3rd leaf, reduced photosynthetic activity, inhibition in overall seedling growth, and failure to develop new (5th) leaf, when subjected to salt stress by transferring them to YS containing 130 mM NaCl for 4 days. As leaf response to salt stress is least investigated in rice by proteomics, we used the 3rd leaf as source material. A comparison of 2-DE protein profiles between the untreated control and salt-stressed 3rd leaves revealed 55 differentially expressed CBB-stained spots, where 47 spots were increased over the control. Of these changed spots, the identity of 33 protein spots (27 increased and 5 decreased) was determined by nESI-LC-MS/MS. Most of these identified proteins belonged to major metabolic processes like photosynthetic carbon dioxide assimilation and photorespiration, suggesting a good correlation between salt stress-responsive proteins and leaf morphology. Moreover, 2-DE immunoblot and enzymatic activity analyses of 3rd leaves revealed remarkable changes in the key marker enzymes associated with oxidative damage to salt stress: ascorbate peroxidase and lipid peroxidation were induced, and catalase was suppressed. These results demonstrate that hydroponic culture system is best suited for proteomics of salt stress in rice seedling.     

Changes of antioxidative enzymes and cell membrane osmosis in tomato colonized by arbuscular Mycorrhizae under NaCl stress

Salinity toxicity is a worldwide agricultural and eco-environmental problem. Many literatures show that arbuscular mycorrhizal fungi (AMF) can enhance salt tolerance of many plants and some physiological changes occurred in AM symbiosis under salt stress. However, the role of ROS-scavenging enzymes in AM tomato is still unknown in continuous salt stress. This study investigated the effect of Glomus mosseae on tomato growth, cell membrane osmosis and examined the antioxidants (superoxide-dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX; peroxidase, POD) responses in roots of mycorrhizal tomato and control under different NaCl stress for 40 days in potted culture. NaCl solution (0, 0.5 and 1%) was added to organic soil in the irrigation water after 45 days inoculated by AMF (Glomus mosseae). (1) AMF inoculation improved tomato growth under salt or saltless condition and reduced cell membrane osmosis, MDA (malonaldehyde) content in salinity. So the salt tolerance of tomato was enhanced by AMF; (2) SOD, APX and POD activity in roots of AM symbiosis were significantly higher than corresponding non-AM plants in salinity or saltless condition. However, CAT activity was transiently induced by AMF and then suppressed to a level similar with non-AM seedlings; (3) higher salinity (1% level) and long stress time suppressed the effect of AMF on SOD, APX, POD and CAT activity; (4) this research suggested that the enhanced salt tolerance in AM symbiosis was mainly related with the elevated SOD, POD and APX activity by AMF which degraded more reactive oxygen species and so alleviated the cell membrane damages under salt stress. Whereas, the elevated SOD, POD and APX activity due to AMF depended on salinity environment.     

Grafted block complex coacervate core micelles and their effect on protein adsorption on silica and polystyrene

We have studied the formation and the stability of grafted block complex coacervate core micelles (C3Ms) in solution and the influence of grafted block C3M coatings on the adsorption of the proteins β-lactoglobulin, bovine serum albumin, and lysozyme. The C3Ms consist of a grafted block copolymer PAA₂₁-b-PAPEO₁₄ (poly(acrylic acid)-b-poly(acrylate methoxy poly(ethylene oxide)), with a negatively charged PAA block and a neutral PAPEO block and a positively charged homopolymer P2MVPI (poly(N-methyl 2-vinyl pyridinium iodide). In solution, these C3Ms partly disintegrate at salt concentrations between 50 and 100 mM NaCl. Adsorption of C3Ms and proteins has been studied with fixed-angle optical reflectometry, at salt concentrations ranging from 1 to 100 mM NaCl. In comparison with the adsorption of PAA₂₁-b-PAPEO₁₄ alone adsorption of C3Ms significantly increases the amount of PAA₂₁-b-PAPEO₁₄ on the surface. This results in a higher surface density of PEO chains. The stability of the C3M coatings and their influence on protein adsorption are determined by the composition and the stability of the C3Ms in solution. A C3M-PAPEO₁₄/P2MVPI₄₃ coating strongly suppresses the adsorption of all proteins on silica and polystyrene. The reduction of protein adsorption is the highest at 100 mM NaCl (>90%). The adsorbed C3M-PAPEO₁₄/P2MVPI₄₃ layer is partly removed from the surface upon exposure to an excess of β-lactoglobulin solution, due to formation of soluble aggregates consisting of β-lactoglobulin and P2MVPI₄₃. In contrast, C3M-PAPEO₁₄/P2MVPI₂₂₈ which has a fivefold longer cationic block enhances adsorption of the negatively charged proteins on both surfaces at salt concentrations above 1 mM NaCl. A single PAA₂₁-b-PAPEO₁₄ layer causes only a moderate reduction of protein adsorption.     

Endophytic Fungi Produce Gibberellins and Indoleacetic Acid and Promotes Host-Plant Growth during Stress

We isolated and examined two endophytic fungi for their potential to secrete phytohormones viz. gibberellins (GAs) and indoleacetic acid (IAA) and mitigate abiotic stresses like salinity and drought. The endophytic fungi Phoma glomerata LWL2 and Penicillium sp. LWL3 significantly promoted the shoot and allied growth attributes of GAs-deficient dwarf mutant Waito-C and Dongjin-beyo rice. Analysis of the pure cultures of these endophytic fungi showed biologically active GAs (GA₁, GA₃, GA₄ and GA₇) in various quantities. The cultures of P. glomerata and Penicillium sp. also contained IAA. The culture application and endophytic-association with host-cucumber plants significantly increased the plant biomass and related growth parameters under sodium chloride and polyethylene glycol induced salinity and drought stress as compared to control plants. The endophytic symbiosis resulted in significantly higher assimilation of essential nutrients like potassium, calcium and magnesium as compared to control plants during salinity stress. Endophytic-association reduced the sodium toxicity and promoted the host-benefit ratio in cucumber plants as compared to non-inoculated control plants. The symbiotic-association mitigated stress by compromising the activities of reduced glutathione, catalase, peroxidase and polyphenol oxidase. Under stress conditions, the endophyte-infection significantly modulated stress through down-regulated abscisic acid, altered jasmonic acid, and elevated salicylic acid contents as compared to control. In conclusion, the two endophytes significantly reprogrammed the growth of host plants during stress conditions.     

The protective role of melatonin under heavy metal-induced stress in Melissa Officinalis L.

Heavy metals, due to their inability to degrade, pose a serious environmental and nutritional problem. The accumulation of essential and non-essential heavy metals in living organisms reduces normal growth and development, resulting in acute poisoning, disease and even death of organisms. Melatonin is a very important multifunctional molecule in protecting plants from oxidative stress due to its ability to directly neutralize reactive oxygen species (ROS). Also, melatonin has a chelating property, which may contribute in reducing metal-induced toxicity. In this paper, the protective role of melatonin in counteracting metal-induced free radical generation was highlighted. Using the HPLC-FLD technique melatonin was identified and quantified in the roots and leaves of lemon balm ( Melissa officinalis L.), grown under photoperiod conditions. Furthermore, the response of plants pre-treated with exogenous 0.1 mM melatonin to the increased zinc (Zn) and cadmium (Cd) concentrations was observed, with changes in mineral (Ca, Mg), physiological and antioxidant status of the plant during heavy metals stress. The obtained melatonin concentrations were the highest published for dry plants so far. Elevated Cd and Zn levels in soil caused alternation in biochemical and physiological parameters of lemon balm leaves and roots. However, melatonin pre-treatment increased plant tolerance to heavy metals stress. Increased Cd and Zn uptake and their translocation into the leaves were also improved, indicating the possible use of melatonin in phytoremediation.