Conferring Salinity Tolerance Research Articles

Over Expression of the Carbonic Anhydrase Gene Confers Salinity Tolerance with Improved Yield in Rice

Background: Abiotic and biotic stresses impact agricultural productivity. Rice being the stable food of India; requires more scientific intervention to boost productivity. Methods: Cloning and transformation of carbonic anhydrase (CA) gene in IR64 rice, molecular analysis of T1 transgenic CA rice plants, salinity tolerance index, leaf disk senescence assay, chlorophyll content, biochemical analysis of antioxidant activities of marker-free CA transgenic lines and measurement of photosynthetic activities and agronomic characteristics of CA transgenic plants were conducted. Result: We have raised two (2) lines of marker free CA overexpressing transgenic rice (Oryza sativa L. cv. IR64) plants. The overexpression of CA driven by CaMV35S promoter in transgenic rice confers high salinity (200 mM NaCl) stress tolerance in the rice. Photosynthetic characteristics such as net photosynthetic rate (PN), stomatal conductance (gs), intercellular CO2 (Ci), as well as chlorophyll (Chl) contents were significantly (22-31%) higher in transgenic lines under salinity stress as compared to control plants. Ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR) and guaiacol peroxidase (GPX) were significantly higher (21-37%) in transgenics as compared with VC and WT plants. Overall, the present research is focused to develop marker free CA overexpressing transgenic lines for better tolerance against salinity stress, higher photosynthesis and better productivity.

Estimates of genetic variability and interplay of germination and seedling traits conferring salinity tolerance in rice (Oryza sativa L.)

This study estimates genetic variability and correlations among germination and seedling traits conferring salinity tolerance in rice accessions. Five rice accessions were screened under salinity levels of 0, 50, 100, and 200 mM NaCl in a controlled laboratory setting. Traits such as germination energy, capacity, shoot and root length, and biomass were measured. Data were analyzed for variance and correlations to assess variability and trait relationships. Significant genetic variability was found among accessions for all traits. Germination energy showed the highest coefficient of variation (CV) at 22.29% under control conditions, while fresh shoot weight had the highest CV (34.35%) under 200 mM salinity. Accessions ACC2 and ACC5 consistently demonstrated higher performance in germination energy (23.33 to 53.33% and 10.00 to 41.67%), germination capacity (40.00 to 60.00% and 28.33 to 46.67%), and shoot length (0.67 to 2.97 cm and 0.40 to 3.93 cm) under various salinity stress levels. ACC1, ACC3, and ACC4 showed more variability but maintained some consistency in specific traits, with ACC4 generally showing lower performance across most traits. Genetic parameter estimates indicated high heritability (˃60%) for all traits, with the highest in germination capacity (96.88%). High genetic advance (GAM) was observed for all traits (˃20%), with germination energy showing the highest (107.00%). Traits with high heritability and genetic advance, such as germination energy, germination capacity, and root length, suggest strong genetic control and potential for improvement through selective breeding. Significant correlations were found between germination energy and capacity (r= 0.89 to 0.96) and between shoot length and leaf length (r= 0.92) under stress conditions. Stress tolerance indices identified accessions ACC2 and ACC5 as the most tolerant, with ACC1 showing consistent performance across traits. This study underscores the importance of identifying resilient traits and accessions to enhance salinity tolerance in rice, contributing to improved productivity in saline-affected regions.

Chitosan nanoparticles (CSNPs) conferred salinity tolerance in maize by upregulating E3 ubiquitin-protein ligase, P5CS1, HKT1, NHX1, and PMP3 genes.

This study explored the transcriptional behaviors of several candidate genes in response to the application of CSNPs (50 and 100 mgl-1) in maize seedlings grown under two salinity levels (NaCl of 0.07 and 0.14 gkg-1soil). Employing CSNPs at both concentrations mitigated the inhibitory role of salinity on the leaf and root fresh weights. The application of CSNPs enhanced the transcription of the E3 ubiquitin-protein ligase gene by an average of threefold, contrasted with the salinity controls. The Δ1-pyrroline-5-carboxylate synthetase (P5CS1) gene was upregulated in response to both individual and mixed treatments of CSNPs and salinity. The transcription of the high-affinity K+ transporter (HKT1) gene displayed an upward trend in response to the CSNPs and salinity treatments. The Na+/H+ exchangers (NHX1) gene exhibited a similar trend to that of the HKT1 gene. The utilization of CSNPs was accompanied by an upregulation in the plasma membrane proteolipid 3 (PMP3) gene, contrasted with the salinity controls. The phenylalanine ammonia-lyase (PAL) activity displayed an upward trend in response to the foliar application of CSNPs. The CSNPs at the 100 mgl-1 concentration were more capable of inducing the ascorbate peroxidase enzyme under both salinity conditions than the 50 mgl-1 dose. The simultaneous exposure of maize seedlings to CSNPs and salinity resulted in the drastic upregulation of the catalase activities. This study provides novel insights into the major mechanisms underlying the stress-mitigating effects of CSNPs, thereby providing a suitable platform for their application in sustainable agricultural practices.

ABA Confers Salinity Tolerance in Lily Cultivars by Modulating Casparian Strip Development, Growth and Physiological Status

ABA Confers Salinity Tolerance in Lily Cultivars by Modulating Casparian Strip Development, Growth and Physiological Status

Synergistic effect of Pseudomonas putida and endomycorrhizal inoculation on the physiological response of onion (Allium cepa L.) to saline conditions

Salinity stress negatively affects the growth and yield of crops worldwide. Onion (Allium cepa L.) is moderately sensitive to salinity. Beneficial microorganisms can potentially confer salinity tolerance. This study investigated the effects of endomycorrhizal fungi (M), Pseudomonas putida (Ps) and their combination (MPs) on onion growth under control (0 ppm), moderate (2000 ppm) and high (4000 ppm) NaCl salinity levels. A pot experiment was conducted with sandy loam soil and onion cultivar Giza 20. Results showed that salinity reduced growth attributes, leaf pigments, biomass and bulb yield while increasing oxidative stress markers. However, individual or combined inoculations significantly increased plant height, bulb diameter and biomass production compared to uninoculated plants under saline conditions. MPs treatment provided the highest stimulation, followed by Pseudomonas and mycorrhizae alone. Overall, dual microbial inoculation showed synergistic interaction, conferring maximum benefits for onion growth, bulbing through integrated physiological and biochemical processes under salinity. Bulb yield showed 3.5, 36 and 83% increase over control at 0, 2000 and 4000 ppm salinity, respectively. In conclusion, combined application of mycorrhizal-Pseudomonas inoculations (MPs) effectively mitigate salinity stress. This approach serves as a promising biotechnology for ensuring sustainable onion productivity under saline conditions.

Relative Expression of a Salinity Stress‐Responsive Na+/H+ Exchanger (NHX) in Root and Leaf Tissues of the African Leafy Vegetable, Amaranthus dubius

ABSTRACTAmaranthus dubius, an African leafy vegetable (ALV), is an easy‐to‐grow, annual shrub and a highly nutritious food source, containing elevated levels of essential nutrients in the leaves. Many ALVs, including A. dubius, can tolerate salinity stress, enabling their cultivation on marginal land. However, the widespread propagation of A. dubius as a stable food source has thus far not been realised due partially to the high frequency at which hybridisation occurs, resulting in high genotypic and phenotypic variability. Therefore, to increase the agricultural output capacity of this species on salt‐affected marginal lands, it is important to screen, select and then clonally propagate the identified salinity‐tolerant genotypes to ensure true‐to‐type fidelity in the regenerated population. It is also important, thereafter, to elucidate their underlying gene expression of the stress response. The present study exposed 4‐week‐old A. dubius seedlings to 100, 200 and 400 mM NaCl to determine their degree of salt tolerance. Genotypes were then screened, selected and clonally propagated through cuttings, based on high growth rates and biomass, and salt tolerance. Generally, growth and physiological parameters decreased as substrate salinity increased. However, individual salt‐stressed genotypes demonstrated similar vigour to nonstressed plants and were able to maintain total protein and chlorophyll concentrations despite increasing salinity. The relative expression of an NHX1‐like transcript was quantified in 15 genotypes using degenerately primed real‐time qPCR. The relative expression of the putative NHX1 gene was 6.7 times greater in root tissues of seedlings treated with 400 mM NaCl (10.7 ± 1.8) compared to the roots of untreated seedlings (1.6 ± 1.3), and 2.8‐fold more than leaf tissues harvested from seedlings treated with 400 mM NaCl. Furthermore, the relative electrical conductivity (EC) of root tissues was 10 times greater than the EC of leaf tissues from the same 400 mM NaCl treatment. Numerous genotypes yielded similar chlorophyll content between 200 and 400 mM NaCl treatments, with genotypes salinity‐1 (S1) (3.5 ± 0.2 μg/cm2) and S34 (4.0 ± 0.4 μg/cm2) having the highest concentrations of chlorophyll in the 400 mM group, which was positively correlated with total protein content. Following micropropagation through direct organogenesis, selected clones maintained true‐to‐type traits such as similar chlorophyll, protein and NHX1‐like expression as their parent plants when exposed to 400 mM NaCl. This study revealed that some genotypes demonstrated salt stress tolerance capabilities rivalling established halophytes by regulating the constitutive or inducible expression of an NHX1‐like protein in roots and leaves. The correlation between protein content and NHX1‐like expression was nonlinear and nonproportional, demonstrating the complexity of this response and necessitating further exploration of specific protein families or functional groups conferring salinity tolerance in this species.

Variation in TaSPL6-D confers salinity tolerance in bread wheat by activating TaHKT1;5-D while preserving yield-related traits.

Na+ exclusion from above-ground tissues via the Na+-selective transporter HKT1;5 is a major salt-tolerance mechanism in crops. Using the expression genome-wide association study and yeast-one-hybrid screening, we identified TaSPL6-D, a transcriptional suppressor of TaHKT1;5-D in bread wheat. SPL6 also targeted HKT1;5 in rice and Brachypodium. A 47-bp insertion in the first exon of TaSPL6-D resulted in a truncated peptide, TaSPL6-DIn, disrupting TaHKT1;5-D repression exhibited by TaSPL6-DDel. Overexpressing TaSPL6-DDel, but not TaSPL6-DIn, led to inhibited TaHKT1;5-D expression and increased salt sensitivity. Knockout of TaSPL6-DDel in two wheat genotypes enhanced salinity tolerance, which was attenuated by a further TaHKT1;5-D knockdown. Spike development was preserved in Taspl6-dd mutants but not in Taspl6-aabbdd mutants. TaSPL6-DIn was mainly present in landraces, and molecular-assisted introduction of TaSPL6-DIn from a landrace into a leading wheat cultivar successfully improved yield on saline soils. The SPL6-HKT1;5 module offers a target for the molecular breeding of salt-tolerant crops.

Oil Palm AP2 Subfamily Gene EgAP2.25 Improves Salt Stress Tolerance in Transgenic Tobacco Plants.

AP2/ERF transcription factor genes play an important role in regulating the responses of plants to various abiotic stresses, such as cold, drought, high salinity, and high temperature. However, less is known about the function of oil palm AP2/ERF genes. We previously obtained 172 AP2/ERF genes of oil palm and found that the expression of EgAP2.25 was significantly up-regulated under salinity, cold, or drought stress conditions. In the present study, the sequence characterization and expression analysis for EgAP2.25 were conducted, showing that it was transiently over-expressed in Nicotiana tabacum L. The results indicated that transgenic tobacco plants over-expressing EgAP2.25 could have a stronger tolerance to salinity stress than wild-type tobacco plants. Compared with wild-type plants, the over-expression lines showed a significantly higher germination rate, better plant growth, and less chlorophyll damage. In addition, the improved salinity tolerance of EgAP2.25 transgenic plants was mainly attributed to higher antioxidant enzyme activities, increased proline and soluble sugar content, reduced H2O2 production, and lower MDA accumulation. Furthermore, several stress-related marker genes, including NtSOD, NtPOD, NtCAT, NtERD10B, NtDREB2B, NtERD10C, and NtP5CS, were significantly up-regulated in EgAP2.25 transgenic tobacco plants subjected to salinity stress. Overall, over-expression of the EgAP2.25 gene significantly enhanced salinity stress tolerance in transgenic tobacco plants. This study lays a foundation for further exploration of the regulatory mechanism of the EgAP2.25 gene in conferring salinity tolerance in oil palm.

Identification of potential morpho-biochemical determinants conferring salt tolerance in wheat (Triticum aestivum L.) through seedling- and -reproductive stage phenotyping

To optimize a reproductive-stage-specific phenotyping protocol and isolate potential determinants conferring salinity tolerance in wheat, two consecutive experiments were conducted using salt-tolerant varieties (Binagom-1 and BARI Gom 25) and a sensitive variety (BARI Gom 20). In the first experiment, seedlings were grown hydroponically, and 14-day-old seedlings were subjected to two different levels of salt stress (EC=12 dS/m and 16 dS/m) for 7 days. Based on the results of tolerant and susceptible varieties, parameters such as root and shoot weight, shoot Na+/K+ ratio, chlorophyll content, proline content, methylglyoxal content, H2O2 content and lipid peroxidation content and the activities of enzymes such as ascorbate peroxidase, peroxidase, and glyoxalase I were considered as potential determinants of salt tolerance. The second experiment employed four leaf cutting treatments under both control and salinity stress conditions. Seedlings were grown in perforated pots filled with field soil, and at the heading stage, plants were subjected to salt stress (12 dS/m) after trimming as indicated. The combined analysis of control and salt stress data obtained from setup B reflected a significant decrease in yield and yield-attributing traits; however, a lesser decrease was observed in tolerant varieties. Correlation studies revealed that grain yield per spike exhibited a significant positive correlation with the number of seeds per spike, spike weight, plant height, and days to first flowering under both stress and control conditions. Additionally, different stress tolerance indices also supported the results of reproductive stage phenotyping. However, further studies will be required to tag the genes/QTLs controlling salinity tolerance in wheat at various growth phases.

Arabidopsis class II TPS controls root development and confers salt stress tolerance through enhanced hydrophobic barrier deposition.

The mechanism of conferring salt tolerance by AtTPS9 involves enhanced deposition of suberin lamellae in the Arabidopsis root endodermis, resulting in reduction of Na+ transported to the leaves. Members of the class I trehalose-6-phosphate synthase (TPS) enzymes are known to play an important role in plant growth and development in Arabidopsis. However, class II TPSs and their functions in salinity stress tolerance are not well studied. We characterized the function of a class II TPS gene, AtTPS9, to understand its role in salt stress response and root development in Arabidopsis. The attps9 mutant exhibited significant reduction of soluble sugar levels in the leaves and formation of suberin lamellae (SL) in the endodermis of roots compared to the wild type (WT). The reduction in SL deposition (hydrophobic barriers) leads to increased apoplastic xylem loading, resulting in enhanced Na+ content in the plants, which explains salt sensitivity of the mutant plants. Conversely, AtTPS9 overexpression lines exhibited increased SL deposition in the root endodermis along with increased salt tolerance, showing that regulation of SL deposition is one of the mechanisms of action of AtTPS9 in conferring salt tolerance to Arabidopsis plants. Our data showed that besides salt tolerance, AtTPS9 also regulates seed germination and root development. qRT-PCR analyses showed significant downregulation of selected SNF1-RELATED PROTEIN KINASE2 genes (SnRK2s) and ABA-responsive genes in the mutant, suggesting that AtTPS9 may regulate the ABA-signaling intermediates as part of the mechanism conferring salinity tolerance.

Effect of salinity stress on growth, chlorophyll, antioxidant enzymes and nutrient content in Azolla spp.

Azolla is an aquatic fern that has a symbiotic association with nitrogen-fixing cyanobacteria. It is mainly used as a biofertilizer in rice; however, its potential under salt-affected rice cultivated area was compromised. Therefore, the present study was undertaken to understand the effect of salinity stress on morpho-physiological, biochemical characteristics, photosynthetic efficacy, nutrient and High Affinity Potassium Transporter (HKT) genes in Azolla. The results indicated that out of 102, 8 Azolla (A. microphylla, BLCC 5, BLCC 18, BLCC 28, Pa Car WTY, R 18, R 54 and R 59) were found tolerant to 80 mM NaCl. The best species for salt tolerant (80 mM NaCl) was A. microphylla, whereas the least-tolerant was A. rubra. Fresh biomass production, frond length and width in A. microphylla were significantly (p < 0.05) higher in A. microphylla than A. rubra in both 40 and 80 mM NaCl. Moreover, chlorophyll a/b ratio, carotenoids and chlorophyll fluorescence (CHF)-derived FO, Fm, Fv/Fm and root architecture (root length, average root diameter, root volume, projectile and surface area) were higher in A. microphylla than A. rubra under 40 and 80 mM NaCl. Contents of Na+ and Ca2+ increased in both A. microphylla and A. rubra, which can interfere with the uptake of essential macronutrients; however, these were accumulated comparatively less in A. microphylla than A. rubra, whereas a reverse trend was observed in cellular accumulation of K+ content. A. microphylla had higher superoxide dismutase (SOD), ascorbate peroxidase (APX), and proline activities in 40 and 80 mM NaCl than A. rubra. For the first time, twenty six HKT primers were designed as a molecular marker to identify salt-tolerant Azolla. Out of these, three HKT primers (Req 6, Aeq14, and Aeq16) were amplified in A. microphylla under NaCl stress, while their amplifications were not observed in A. rubra (salt susceptible). In A. microphylla, the expression of the Req 6 (HKT) gene were more under NaCl stress. Moreover, further research is needed to discover and validate the biochemical and molecular processes that confer salinity tolerance in Azolla plants.

Low apoplastic Na+ and intracellular ionic homeostasis confer salinity tolerance upon Ca2SiO4 chemigation in Zea mays L. under salt stress.

Salinity is known to have a greater impact on shoot growth than root growth. Na+ buildup in plant tissue under salt stress has been proposed as one of the main issues that causes growth inhibition in crops via ionic imbalances, osmotic stress and pH disturbances. However, the evidence for apoplastic Na+ buildup and the role of silicon in Na+ accumulation at the subcellular level is still enigmatic. The current study focuses on the accumulation of Na+ in the apoplast and symplast of younger and older leaves of two maize varieties (Iqbal as salt-tolerant and Jalal as salt-sensitive) using hydroponic culture along with silicon supplementation under short-term salinity stress. Subcellular ion analysis indicated that silicon nutrition decreased Na+ concentration in both apoplastic washing fluid and symplastic fluid of maize under salt stress. The addition of silicon under NaCl treatment resulted in considerable improvement in fresh biomass, relative water content, chlorophyll content, and concentration of important subcellular ions (i.e., Ca2+, Mg2+, and K+). Knowledge of subcellular ion analysis is essential for solving the mechanisms underlying vital cellular functions e.g. in the current study, the soluble Na+ concentration in the apoplast of older leaves was found to be significantly greater (36.1 mM) in the salt-sensitive variety under NaCl treatment, which was 42.4% higher when compared to the Na+ concentration in the salt-tolerant variety under the same treatment which can influence permeability of cell membrane, signal transduction pathways and provides insights into how ion compartmentalization can contributes to salt tolerance. Calcium silicate enrichment can contribute to increased growth and improved ionic homeostasis by minimizing leaf electrolyte leakage, improving mechanical functions of cell wall and reducing water loss, and improved photosynthetic function. In current investigation, increased water content and intracellular ionic homeostasis along with reduced concentration of Na+ in the maize leaf apoplast suggest that calcium silicate can be used to ameliorate the adverse effects of salt stress and obtain yield using marginal saline lands.

One size does not fit all: Different strategies employed by triticale and barley plants to deal with soil salinity

Salinity stress tolerance is a polygenic trait conferred by multiple physiological mechanisms, operating at various levels of plant structural organization. In this work, we have compared adaptative strategies employed by barley and triticale, two most tolerant cereal species, to deal with saline conditions. While the photosynthetic attributes were considerably reduced by 200 mM NaCl treatment in both species, triticale plants maintained higher photosynthetic capacity per unit leaf area compared with barley. Triticale plants also showed higher stomatal conductance under both saline and non-saline conditions, explaining the above photosynthetic rate. However, this high photosynthetic capacity in triticale was not translated into higher biomass (as compared with barley) due to stronger salinity-induced inhibition of tillering in the former species. Leaf elemental analysis revealed that salinity tolerance in triticale was related to its ability to exclude Na+ from the shoot while tolerant barley plant tended to accumulate more Na in shoot upon exposure to salt stress, for osmotic adjustment purposes. Salinity tolerance in both species was also causally associated with higher Fe content that was essential for activation of superoxide dismutase (SOD) to reduce the extent of oxidative stress. Electrophysiological experiments also revealed several patterns associated with control of ion transport in root epidermis, of which desensitization of K+- and Ca2+- ROS-inducible cation channels were the main trait conferring salinity tolerance. Cytosolic K+ retention in the root elongation zone in response to salt stress was more efficient in both triticale genotypes compared to barley. Finally, barley and salt tolerant genotypes of triticale showed higher speed of stomata, thus possessing higher water use efficiency.

Reactive oxygen species (ROS) scavenging and methylglyoxal (MG) detoxification confers salinity tolerance in mustard cultivars inoculated with Piriformospora (Serendipita) indica

Salinization has become a major concern for the growth and development of plants, resulting in significant crop losses. Beneficial microbial symbionts are utilized as promising tools in alleviation of adverse effects of salt stress. Piriformospora (Serendipita) indica is one of the cost-effective, eco-friendly, and stress relieving biostimulant that positively regulates the growth of a broad host range of plants. The present study was to investigate the impact of P. indica on Indian mustard (Brassica juncea L.) cultivars (Pusa Jaikisan and Pusa Bold) under different levels of salinity stress (0, 50, 100, 150, 200 and 300 mM NaCl) by analyzing morphological, physiological and biochemical attributes. Results revealed that the plant biomass, chlorophyll content, and relative water content were reduced and Na+ and Cl‒ ions uptake increased with increasing NaCl concentrations. The maximum reduction in the measured traits was observed at 300 mM NaCl in both cultivars [tolerant (Pusa Jaikisan) and sensitive (Pusa Bold)]. Additionally, the oxidative stress markers (TBARS, MG, H2O2, and EL) accumulation were increased with increasing salt concentrations. However, P. indica colonization enhanced the plant performance by improving the biomass, chlorophyll, and water content, and reduced levels of oxidative stress markers (TBARS, MG, H2O2, and EL) in both cultivars comparatively more in Pusa Jaikisan than Pusa Bold under salinity stress. Additionally, the maximum activities of methyl glyoxal detoxifying enzymes (Gly I & Gly II) along with the components of ROS scavenging machinery (SOD, APX, GPX, GR, AsA, and GSH) in the P. indica colonized plants conferred salt tolerance in comparison to non-colonized cultivars under salinity stress. The results indicate that P. indica promotes the growth and tolerance of mustard plants under salinity stress, thus providing opportunity for further assessment in terms of use in sustainable agriculture.

Stress-responsive gene regulation conferring salinity tolerance in wheat inoculated with ACC deaminase producing facultative methylotrophic actinobacterium.

Microbes enhance crop resilience to abiotic stresses, aiding agricultural sustainability amid rising global land salinity. While microbes have proven effective via seed priming, soil amendments, and foliar sprays in diverse crops, their mechanisms remain less explored. This study explores the utilization of ACC deaminase-producing Nocardioides sp. to enhance wheat growth in saline environments and the molecular mechanisms underlying Nocardioides sp.-mediated salinity tolerance in wheat. The Nocardioides sp. inoculated seeds were grown under four salinity regimes viz., 0 dS m-1, 5 dS m-1, 10 dS m-1, and 15 dS m-1, and vegetative growth parameters including shoot-root length, germination percentage, seedling vigor index, total biomass, and shoot-root ratio were recorded. The Nocardioides inoculated wheat plants performed well under saline conditions compared to uninoculated plants and exhibited lower shoot:root (S:R) ratio (1.52 ± 0.14 for treated plants against 1.84 ± 0.08 for untreated plants) at salinity level of 15 dS m-1 and also showed improved biomass at 5 dS m-1 and 10 dS m-1. Furthermore, the inoculated plants also exhibited higher protein content viz., 22.13 mg g-1, 22.10 mg g-1, 22.63 mg g-1, and 23.62 mg g-1 fresh weight, respectively, at 0 dS m-1, 5 dS m-1, 10 dS m-1, and 15 dS m-1. The mechanisms were studied in terms of catalase, peroxidase, superoxide dismutase, and ascorbate peroxidase activity, free radical scavenging potential, in-situ localization of H2O2 and superoxide ions, and DNA damage. The inoculated seedlings maintained higher enzymatic and non-enzymatic antioxidant potential, which corroborated with reduced H2O2 and superoxide localization within the tissue. The gene expression profiles of 18 stress-related genes involving abscisic acid signaling, salt overly sensitive (SOS response), ion transporters, stress-related transcription factors, and antioxidant enzymes were also analyzed. Higher levels of stress-responsive gene transcripts, for instance, TaABARE (~+7- and +10-fold at 10 dS m-1 and 15 dS m-1); TaHAk1 and hkt1 (~+4- and +8-fold at 15 dS m-1); antioxidant enzymes CAT, MnSOD, POD, APX, GPX, and GR (~+4, +3, +5, +4, +9, and +8 folds and), indicated actively elevated combat mechanisms in inoculated seedlings. Our findings emphasize Nocardioides sp.-mediated wheat salinity tolerance via ABA-dependent cascade and salt-responsive ion transport system. This urges additional study of methylotrophic microbes to enhance crop abiotic stress resilience.

Synergistic Effects of Rhizobacteria and Salicylic Acid on Maize Salt-Stress Tolerance.

Maize (Zea mays L.) is a salt-sensitive plant that experiences stunted growth and development during early seedling stages under salt stress. Salicylic acid (SA) is a major growth hormone that has been observed to induce resistance in plants against different abiotic stresses. Furthermore, plant growth-promoting rhizobacteria (PGPR) have shown considerable potential in conferring salinity tolerance to crops via facilitating growth promotion, yield improvement, and regulation of various physiological processes. In this regard, combined application of PGPR and SA can have wide applicability in supporting plant growth under salt stress. We investigated the impact of salinity on the growth and yield attributes of maize and explored the combined role of PGPR and SA in mitigating the effect of salt stress. Three different levels of salinity were developed (original, 4 and 8 dS m-1) in pots using NaCl. Maize seeds were inoculated with salt-tolerant Pseudomonas aeruginosa strain, whereas foliar application of SA was given at the three-leaf stage. We observed that salinity stress adversely affected maize growth, yield, and physiological attributes compared to the control. However, both individual and combined applications of PGPR and SA alleviated the negative effects of salinity and improved all the measured plant attributes. The response of PGPR + SA was significant in enhancing the shoot and root dry weights (41 and 56%), relative water contents (32%), chlorophyll a and b contents (25 and 27%), and grain yield (41%) of maize under higher salinity level (i.e., 8 dS m-1) as compared to untreated unstressed control. Moreover, significant alterations in ascorbate peroxidase (53%), catalase (47%), superoxide dismutase (21%), MDA contents (40%), Na+ (25%), and K+ (30%) concentration of leaves were pragmatic under combined application of PGPR and SA. We concluded that integration of PGPR and SA can efficiently induce salinity tolerance and improve plant growth under stressed conditions.

Exogenous Application of Moringa Leaf Extract Confers Salinity Tolerance in Sunflower by Concerted Regulation of Antioxidants and Secondary Metabolites

Exogenous Application of Moringa Leaf Extract Confers Salinity Tolerance in Sunflower by Concerted Regulation of Antioxidants and Secondary Metabolites

AtCIPK16, a CBL-interacting protein kinase gene, confers salinity tolerance in transgenic wheat.

Globally, wheat is the major source of staple food, protein, and basic calories for most of the human population. Strategies must be adopted for sustainable wheat crop production to fill the ever-increasing food demand. Salinity is one of the major abiotic stresses involved in plant growth retardation and grain yield reduction. In plants, calcineurin-B-like proteins form a complicated network with the target kinase CBL-interacting protein kinases (CIPKs) in response to intracellular calcium signaling as a consequence of abiotic stresses. The AtCIPK16 gene has been identified in Arabidopsis thaliana and found to be significantly upregulated under salinity stress. In this study, the AtCIPK16 gene was cloned in two different plant expression vectors, i.e., pTOOL37 having a UBI1 promoter and pMDC32 having a 2XCaMV35S constitutive promoter transformed through the Agrobacterium-mediated transformation protocol, in the local wheat cultivar Faisalabad-2008. Based on their ability to tolerate different levels of salt stress (0, 50, 100, and 200 mM), the transgenic wheat lines OE1, OE2, and OE3 expressing AtCIPK16 under the UBI1 promoter and OE5, OE6, and OE7 expressing the same gene under the 2XCaMV35S promoter performed better at 100 mM of salinity stress as compared with the wild type. The AtCIPK16 overexpressing transgenic wheat lines were further investigated for their K+ retention ability in root tissues by utilizing the microelectrode ion flux estimation technique. It has been demonstrated that after 10min of 100 mM NaCl application, more K+ ions were retained in the AtCIPK16 overexpressing transgenic wheat lines than in the wild type. Moreover, it could be concluded that AtCIPK16 functions as a positive elicitor in sequestering Na+ ions into the cell vacuole and retaining more cellular K+ under salt stress to maintain ionic homeostasis.

Genome-Wide Identification and Characterization of the Polyamine Uptake Transporter (Put) Gene Family in Tomatoes and the Role of Put2 in Response to Salt Stress

The polyamine uptake transporter (Put), an important polyamines-related protein, is involved in plant cell growth, developmental processes, and abiotic stimuli, but no research on the Put family has been carried out in the tomato. Herein, eight tomato Put were identified and scattered across four chromosomes, which were classified into three primary groups by phylogenetic analysis. Protein domains and gene structural organization also showed a significant degree of similarity, and the Put genes were significantly induced by various hormones and polyamines. Tissue-specific expression analysis indicated that Put genes were expressed in all tissues of the tomato. The majority of Put genes were induced by different abiotic stresses. Furthermore, Put2 transcription was found to be responsive to salt stress, and overexpression of Put2 in yeast conferred salinity tolerance and polyamine uptake. Moreover, overexpression of Put2 in tomatoes promoted salinity tolerance accompanied by a decrease in the Na+/K+ ratio, restricting the generation of reactive oxygen and increasing polyamine metabolism and catabolism, antioxidant enzyme activity (SOD, CAT, APX, and POD), and nonenzymatic antioxidant activity (GSH/GSSG and ASA/DHA ratios, GABA, and flavonoid content); loss of function of put2 produced opposite effects. These findings highlight that Put2 plays a pivotal role in mediating polyamine synthesis and catabolism, and the antioxidant capacity in tomatoes, providing a valuable gene for salinity tolerance in plants.

Exogenous Gallic Acid Confers Salt Tolerance in Rice Seedlings: Modulation of Ion Homeostasis, Osmoregulation, Antioxidant Defense, and Methylglyoxal Detoxification Systems

The worldwide saline-affected area is expanding day by day, and soil salinity restricts crop development and productivity, including rice. Considering this, the current study explored the response of gallic acid (GA) in conferring salinity tolerance in rice seedlings. Fourteen-day-old rice (Oryza sativa L. cv. BRRI dhan52) seedlings were treated with 200 mM NaCl alone or combined with 1 mM GA. Salt stress resulted in osmotic, ionic, and oxidative stress in rice seedlings. Osmotic stress increased proline accumulation and osmotic potential, which decreased the relative water content, chlorophyll contents, and dry weight. Ionic stress interrupted ion homeostasis by Na+ accumulation and K+ leakage. Osmotic and ionic stress, concomitantly, disrupted antioxidant defense and glyoxalase systems by higher production of reactive oxygen species (ROS) and methylglyoxal (MG), respectively. It resulted in oxidative damage indicated by the high amount of malondialdehyde (MDA). The supplementation of GA in salt-treated rice seedlings partially recovered salt-induced damages by improving osmotic and ionic homeostasis by increasing water balance and decreasing Na+ content and Na+/K+ ratio. Supplemental GA enhanced the antioxidant defense system in salt-treated rice seedlings by increasing ascorbate (AsA), glutathione (GSH), and phenolic compounds and the activities of AsA-GSH cycle enzymes, including monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and glutathione reductase (GR) enzymes that accelerated ROS detoxification and decreased oxidative damage. Gallic acid also enhanced the detoxification of MG by triggering glyoxalase enzyme activities in salt-treated rice seedlings. The present findings elucidated that supplemental GA reversed salt-induced damage in rice seedlings through improving osmotic and ionic homeostasis and upregulating the ROS and MG detoxification system.