Contribution of Antioxidant System Components to the Long-Term Physiological and Protective Effect of Salicylic Acid on Wheat under Salinity Conditions.

Salicylic acid (SA) plays a crucial role in regulating plant growth and development and mitigating the negative effects of various stresses, including salinity. In this study, the effect of 50 µM SA on the physiological and biochemical parameters of wheat plants under normal and stress conditions was investigated. The results showed that on the 28th day of the growing season, SA pretreatment continued to stimulate the growth of wheat plants. This was evident through an increase in shoot length and leaf area, with the regulation of leaf blade width playing a significant role in this effect. Additionally, SA improved photosynthesis by increasing the content of chlorophyll a (Chl a) and carotenoids (Car), resulting in an increased TAP (total amount of pigments) index in the leaves. Furthermore, SA treatment led to a balanced increase in the levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) in the leaves, accompanied by a slight but significant accumulation of ascorbic acid (ASA), hydrogen peroxide (H2O2), proline, and the activation of glutathione reductase (GR) and ascorbate peroxidase (APX). Exposure to salt stress for 28 days resulted in a reduction in length and leaf area, photosynthetic pigments, and GSH and ASA content in wheat leaves. It also led to the accumulation of H2O2 and proline and significant activation of GR and APX. However, SA pretreatment exhibited a long-term growth-stimulating and protective effect under stress conditions. It significantly mitigated the negative impacts of salinity on leaf area, photosynthetic pigments, proline accumulation, lipid peroxidation, and H2O2. Furthermore, SA reduced the salinity-induced depletion of GSH and ASA levels, which was associated with the modulation of GR and APX activities. In small-scale field experiments conducted under natural growing conditions, pre-sowing seed treatment with 50 µM SA improved the main indicators of grain yield and increased the content of essential amino acids in wheat grains. Thus, SA pretreatment can be considered an effective approach for providing prolonged protection to wheat plants under salinity and improving grain yield and quality.

Seed Treatment with Sodium Nitroprusside Ensures a Long-Term Physiological and Protective Effect on Wheat under Salinity.

Although salinity inhibits plant growth, the use of a nitric oxide (NO) gasotransmitter can reduce its negative effects. In this study, the influence of 200 µM sodium nitroprusside (SNP) (donor of NO) on wheat plants (Triticum aestivum L., cv. Salavat Yulaev) in conditions of salinization (100 mM NaCl) was analyzed in pot experiments. Seed priming regulated the level of endogenous NO in normal and salinity conditions throughout the entire experiment (30 and 60 days). Salinity led to the strong accumulation of NO and H2O2, which is negative for plants, and significantly reduced leaf area and photosynthetic pigments (chlorophyll a and b and carotenoids). In addition, stress caused a drop in the content of reduced glutathione (GSH) and ascorbic acid (ASA), an accumulation of oxidized glutathione (GSSG), and significantly activated glutathione reductase (GR), ascorbate peroxidase (APX), and lipid peroxidation (LPO) in wheat leaves. SNP treatment significantly attenuated the negative effects of salinity on leaf area and photosynthetic pigments. An important indicator of reducing the damaging effect of salinity on treated plants is the stabilization of the content of GSH and ASA throughout the experiment (60 days). This condition has been associated with long-term modulation of GR and APX activity. Such an effect of 200 µM SNP may be related to its ability to reduce stress-induced accumulation of NO. Additional accumulation of proline also mitigated the negative effect of salinity on plants, and this also evidenced decreased LPO and H2O2 in them. For the first time, in natural growing conditions (small-scale field experiments), it was found that pre-sowing seed treatment with 200 µM SNP led to an improvement in the main yield indicators and an increase in the content of essential amino acids in wheat grains. Thus, SNP treatment can be used as an effective approach for prolonged protection of wheat plants under salinity and to improve grain yield and its quality.

Endophytic Plant Growth-Promoting Bacterium Bacillus subtilis Reduces the Toxic Effect of Cadmium on Wheat Plants.

Heavy metal ions, in particular cadmium (Cd), have a negative impact on the growth and productivity of major crops, including wheat. The use of environmentally friendly approaches, in particular, bacteria that have a growth-stimulating and protective effect, can increase the resistance of plants. The effects of the pre-sowing seed treatment with the plant growth-promoting endophyte Bacillus subtilis 10-4 (BS) on cadmium acetate (Cd)-stressed Triticum aestivum L. (wheat) growth, photosynthetic pigments, oxidative stress parameters, roots' lignin content, and Cd ions accumulation in plants were analyzed. The results showed that the tested Cd-tolerant BS improved the ability of wheat seeds to germinate in the presence of different Cd concentrations (0, 0.1, 0.5, and 1 mM). In addition, the bacterial treatment significantly decreased the damaging effects of Cd stress (1 mM) on seedlings' linear dimensions (lengths of roots and shoots), biomass, as well as on the integrity and permeability of the cell walls (i.e., lipid peroxidation and electrolyte leakage) and resulted in reduced H2O2 generation. The pretreatment with BS prevented the Cd-induced degradation of the leaf photosynthetic pigments chlorophyll (Chl) a, Chl b, and carotenoids. Moreover, the bacterial treatment intensified the lignin deposition in the roots under normal and, especially, Cd stress conditions, thereby enhancing the barrier properties of the cell wall. This manifested in a reduced Cd ions accumulation in the roots and in the restriction of its translocation to the aboveground parts (shoots) of the bacterized plants under Cd stress in comparison with non-bacterized controls. Thus, the pre-sowing seed treatment with the endophyte BS may serve as an eco-friendly approach to improve wheat production in Cd-contaminated areas.

Components of the Phenylpropanoid Pathway in the Implementation of the Protective Effect of Sodium Nitroprusside on Wheat under Salinity.

Nitric oxide (NO) is a multifunctional, gaseous signaling molecule implicated in both physiological and protective responses to biotic and abiotic stresses, including salinity. In this work, we studied the effects of 200 µM exogenous sodium nitroprusside (SNP, a donor of NO) on the components of the phenylpropanoid pathway, such as lignin and salicylic acid (SA), and its relationship with wheat seedling growth under normal and salinity (2% NaCl) conditions. It was established that exogenous SNP contributed to the accumulation of endogenous SA and increased the level of transcription of the pathogenesis-related protein 1 (PR1) gene. It was found that endogenous SA played an important role in the growth-stimulating effect of SNP, as evidenced by the growth parameters. In addition, under the influence of SNP, the activation of phenylalanine ammonia lyase (PAL), tyrosine ammonia lyase (TAL), and peroxidase (POD), an increase in the level of transcription of the TaPAL and TaPRX genes, and the acceleration of lignin accumulation in the cell walls of roots were revealed. Such an increase in the barrier properties of the cell walls during the period of preadaptation played an important role in protection against salinity stress. Salinity led to significant SA accumulation and lignin deposition in the roots, strong activation of TAL, PAL, and POD, and suppression of seedling growth. Pretreatment with SNP under salinity conditions resulted in additional lignification of the root cell walls, decreased stress-induced endogenous SA generation, and lower PAL, TAL, and POD activities in comparison to untreated stressed plants. Thus, the obtained data suggested that during pretreatment with SNP, phenylpropanoid metabolism was activated (i.e., lignin and SA), which contributed to reducing the negative effects of salinity stress, as evidenced by the improved plant growth parameters.

Effects of Pseudomonas sp. OBA 2.4.1 on Growth and Tolerance to Cadmium Stress in Pisum sativum L.

Cadmium stress is a barrier to crop production, yield, quality, and sustainable agriculture. In the current study, we investigated the characteristics of bacterial strain Pseudomonas sp. OBA 2.4.1 under cadmium (CdCl2) stress and its influence on Cd stresses in pea (Pisum sativum L.) seedlings. It was revealed that strain OBA 2.4.1 is tolerant of up to 2 mM CdCl2, and seed treatment with the bacterium enhanced pea plant growth (length of seedlings) under 0.5 mM cadmium stress. This bacterial strain showed plant growth-promoting properties, including biofilm formation and siderophore activity. An important advantage of the studied strain OBA 2.4.1 is its ability to colonize the plant roots. Moreover, the inoculation with strain OBA 2.4.1 significantly reduced oxidative stress markers in pea seedlings under cadmium stress. These findings suggest that cadmium stress-tolerant strain OBA 2.4.1 could enhance pea plant growth by mitigating stress-caused damage, possibly providing a baseline and eco-friendly approach to address heavy metal stress for sustainable agriculture.

Role of Endogenous Salicylic Acid as a Hormonal Intermediate in the Bacterial Endophyte Bacillus subtilis-Induced Protection of Wheat Genotypes Contrasting in Drought Susceptibility under Dehydration.

Endophytic Bacillus subtilis is a non-pathogenic beneficial bacterium which promotes plant growth and tolerance to abiotic stresses, including drought. However, the underlying physiological mechanisms are not well understood. In this study, the potential role that endogenous salicylic acid (SA) plays in regulating endophytic B. subtilis-mediated drought tolerance in wheat (Triticum aestivum L.) was examined. The study was conducted on genotypes with contrasting levels of intrinsic drought tolerance (drought-tolerant (DT) cv. Ekada70; drought-susceptible (DS) cv. Salavat Yulaev). It was revealed that B. subtilis 10-4 promoted endogenous SA accumulation and increased the relative level of transcripts of the PR-1 gene, a marker of the SA-dependent defense pathway, but two wheat cultivars responded differently, with the highest levels exhibited in DT wheat seedlings. These had a positive correlation with the ability of strain 10-4 to effectively protect DT wheat seedlings against drought injury by decreasing osmotic and oxidative damages (i.e., proline, water holding capacity (WHC), and malondialdehyde (MDA)). However, the use of the SA biosynthesis inhibitor 1-aminobenzotriazole prevented endogenous SA accumulation under normal conditions and the maintenance of its increased level under stress as well as abolished the effects of B. subtilis treatment. Particularly, the suppression of strain 10-4-induced effects on proline and WHC, which are both contributing factors to dehydration tolerance, was found. Moreover, the prevention of strain 10-4-induced wheat tolerance to the adverse impacts of drought, as judged by the degree of membrane lipid peroxidation (MDA) and plant growth (length, biomass), was revealed. Thus, these data provide an argument in favor of a key role of endogenous SA as a hormone intermediate in triggering the defense responses by B. subtilis 10-4, which also afford the foundation for the development of the bacterial-induced tolerance of these two different wheat genotypes under dehydration.

Effects of Rhizobium leguminosarum Thy2 on the Growth and Tolerance to Cadmium Stress of Wheat Plants.

Cadmium (Cd) stress is an obstacle for crop production, quality crops, and sustainable agriculture. An important role is played by the application of eco-friendly approaches to improve plant growth and stress tolerance. In the current study, a pre-sowing seed treatment with Rhizobium leguminosarum strains, isolated from the leguminous plants Phaseolus vulgaris (strain Pvu5), Vicia sylvatica (strain VSy12), Trifolium hybridium (strain Thy2), and T. pratense (strain TPr4), demonstrated different effects on wheat (Triticum aestivum L.) plant growth under normal conditions. Among all tested strains, Thy2 significantly increased seed germination, seedling length, fresh and dry biomass, and leaf chlorophyll (Chl) content. Further analysis showed that Thy2 was capable of producing indole-3-acetic acid and siderophores and fixing nitrogen. Under Cd stress, Thy2 reduced the negative effect of Cd on wheat growth and photosynthesis and had a protective effect on the antioxidant system. This was expressed in the additional accumulation of glutathione and proline and the activation of glutathione reductase. In addition, Thy2 led to a significant reduction in oxidative stress, which was evidenced by the data on the stabilization of the ascorbate content and the activity of ascorbate peroxidase. In addition, Thy2 markedly reduced Cd-induced membrane lipid peroxidation and electrolyte leakage in the plants. Thus, the findings demonstrated the ability of the R. leguminosarum strain Thy2, isolated from T. hybridium nodules, to exert a growth-promoting and anti-stress effect on wheat plants. These results suggest that the Thy2 strain may enhance wheat plant growth by mitigating Cd stress, particularly through improving photosynthesis and antioxidant capacity and reducing the severity of oxidative damage. This may provide a basic and biological approach to use the Thy2 strain as a promising, eco-friendly candidate to combat Cd stress in wheat production.

Contribution of Ascorbate and Glutathione in Endobacteria Bacillus subtilis-Mediated Drought Tolerance in Two Triticum aestivum L. Genotypes Contrasting in Drought Sensitivity.

We evaluated the effect of endobacteria Bacillus subtilis (strain 10-4) as a co-inoculant for promoting plant growth and redox metabolism in two contrasting genotypes of Triticum aestivum L. (wheat): Ekada70 (drought tolerant (DT)) and Salavat Yulaev (drought susceptible (DS)) in early stages of adaptation to drought (12% PEG-6000). Results revealed that drought reduced growth and dramatically augmented oxidative stress markers, i.e., hydrogen peroxide (H2O2) and lipid peroxidation (MDA). Furthermore, the depletion of ascorbate (AsA) and glutathione (GSH), accompanied by a significant activation of ascorbate peroxidase (APX) and glutathione reductase (GR), in both stressed wheat cultivars (which was more pronounced in DS genotype) was found. B. subtilis had a protective effect on growth and antioxidant status, wherein the stabilization of AsA and GSH levels was revealed. This was accompanied by a decrease of drought-caused APX and GR activation in DS plants, while in DT plants additional antioxidant accumulation and GR activation were observed. H2O2 and MDA were considerably reduced in both drought-stressed wheat genotypes because of the application of B. subtilis. Thus, the findings suggest the key roles in B. subtilis-mediated drought tolerance in DS cv. Salavat Yulaev and DT cv. Ekada70 played are AsA and GSH, respectively; which, in both cases, resulted in reduced cell oxidative damage and improved growth in seedlings under drought.

Seed Priming with Endophytic Bacillus subtilis Modulates Physiological Responses of Two Different Triticum aestivum L. Cultivars under Drought Stress.

The protective effects against drought stress of the endophytic bacterium Bacillus subtilis 10-4 were measured by studying the priming response in two wheat (Triticum aestivum L.)-Ekada70 (E70) and Salavat Yulaev (SY)-lines, tolerant and susceptible to drought, respectively. B. subtilis 10-4 improved germination and growth parameters under normal conditions in both cultivars with the most pronounced effect observed in cv. E70. Under drought conditions, B. subtilis 10-4 significantly ameliorated the negative impact of stress on germination and growth of cv. E70, but had no protective effect on cv. SY. B. subtilis 10-4 induced an increase in the levels of photosynthetic chlorophyll (Chl) a, Chl b, and carotenoids (Car) in the leaves of cv. E70, both under normal and drought conditions. In cv. SY plants, bacterial inoculation decreased the contents of Chl a, Chl b, and Car under normal conditions, but pigment content were almost recovered under drought stress. B. subtilis 10-4 increased water holding capacity (WHC) of cv. E70 (but did not affect this parameter in cv. SY) and prevented the stress-induced decline in WHC in both cultivars. Notably, B. subtilis 10-4 increased endogenous salicylic acid (SA) concentration in both cultivars, especially in cv. E70. Moreover, B. subtilis 10-4 reduced drought-induced endogenous SA accumulation, which was correlated with the influence of endophyte on growth, indicating a possible involvement of endogenous SA in the implementation of B. subtilis-mediated effects in both cultivars. Overall, B. subtilis 10-4 inoculation was found to increase drought tolerance in seedlings of both cultivars, as evidenced by decreased lipid peroxidation, proline content, and electrolyte leakage from tissues of wheat seedlings primed with B. subtilis 10-4 under drought conditions.

Wheat germ agglutinin is involved in the protective action of 24-epibrassinolide on the roots of wheat seedlings under drought conditions.

The involvement of wheat germ agglutinin (WGA) in the protective action of 24-epibrassinolide (EBR) against drought stress was studied in the seedling roots of two wheat cultivars differing in drought tolerance. Under dehydration conditions, the contents of ABA and WGA were shown to change significantly in the roots of either drought-tolerant cultivar Omskaya 35 or drought-sensitive cultivar Salavat Yulaev. Meanwhile, accumulation of either ABA or WGA started earlier and was two times greater in plants of drought-tolerant cultivar. Since WGA is an excreted protein, it is not surprising that the level of lectin in the roots gradually decreased by the 7th day of treatment due to its exudation into root environment. Pre-sowing treatment with EBR contributed to additional accumulation of lectin as compared to the control variants of either cultivar, while the hormone treatment did not change ABA content. Meanwhile, under conditions of drought, EBR-pretreated seedlings were characterized by lower level of accumulation of ABA and WGA in the roots. EBR application was found to prevent drought-induced inhibition of cell division in the root apical meristem, while WGA excreted into the root environment may contribute significantly to the effect.

Involvement of dehydrins in 24-epibrassinolide-induced protection of wheat plants against drought stress.

In this study, we performed a comparative analysis of the physiological and biochemical parameters of wheat cultivars with contrasting drought resistance, drought-resistant Omskaya 35 (O-35) and less drought-resistant Salavat Yulaev (SYu), during 7-day germination under drought stress simulated by 5% mannitol. In addition, we evaluated the effectiveness of pre-sowing seed treatment with 0.4 µM 24-epibrassinolide (EBR) used to increase the resistance of plants of both cultivars to drought stress. It was revealed that mannitol has caused significant changes in the hormonal balance of the plants of both cultivars, associated with abscisic acid (ABA) accumulation and decrease in the contents of indoleacetic acid (IAA) and cytokinins (CKs). It should be noted that more dramatic changes in the content of phytohormones were characteristic for seedlings of SYu cultivar, which was reflected in a stronger growth inhibition of these plants. Pretreatment with EBR mitigated the negative effect of drought on the hormonal status and growth of seedlings during their germination. Furthermore, we found that drought caused accumulation of dehydrin (DHN) proteins, especially of low molecular weight DHNs, whose abundance was 2.5 times greater in O-35 cultivar than in SYu plants. EBR-pretreated plants of both cultivars were characterized by the additional accumulation of DHNs, indicating their involvement in the development of the EBR-induced wheat drought resistance. The use of fluridone allowed us to demonstrate ABA-dependent and ABA-independent pathways of regulation of low molecular mass dehydrins accumulation by EBR in wheat plants of both cultivars under drought conditions.