Regulatory Role of Circadian Clocks on ABA Production and Signaling, Stomatal Responses, and Water-Use Efficiency under Water-Deficit Conditions

Plants deploy molecular, physiological, and anatomical adaptations to cope with long-term water-deficit exposure, and some of these processes are controlled by circadian clocks. Circadian clocks are endogenous timekeepers that autonomously modulate biological systems over the course of the day–night cycle. Plants’ responses to water deficiency vary with the time of the day. Opening and closing of stomata, which control water loss from plants, have diurnal responses based on the humidity level in the rhizosphere and the air surrounding the leaves. Abscisic acid (ABA), the main phytohormone modulating the stomatal response to water availability, is regulated by circadian clocks. The molecular mechanism of the plant’s circadian clock for regulating stress responses is composed not only of transcriptional but also posttranscriptional regulatory networks. Despite the importance of regulatory impact of circadian clock systems on ABA production and signaling, which is reflected in stomatal responses and as a consequence influences the drought tolerance response of the plants, the interrelationship between circadian clock, ABA homeostasis, and signaling and water-deficit responses has to date not been clearly described. In this review, we hypothesized that the circadian clock through ABA directs plants to modulate their responses and feedback mechanisms to ensure survival and to enhance their fitness under drought conditions. Different regulatory pathways and challenges in circadian-based rhythms and the possible adaptive advantage through them are also discussed.Peer Reviewe

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

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

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

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

Application of Endophytic Bacillus subtilis and Salicylic Acid to Improve Wheat Growth and Tolerance under Combined Drought and Fusarium Root Rot Stresses

In nature, plants are constantly exposed to a varied abiotic and biotic stresses or their combinations, limiting the productivity of major crops, including wheat. Combinations of drought and soil-borne Fusarium-instigated diseases are the most common combinations of stresses, significantly reducing wheat yield around the world. Here, were analyzed the potential of application of endophytic bacteria Bacillus subtilis (strain 10–4) together with the natural signal molecule salicylic acid (SA) to improve growth and tolerance of Triticum aestivum L. (wheat) plants under combined drought and Fusarium culmorum-instigated root rot (FRR) stresses. It was revealed that pre-sowing treatment with B. subtilis 10–4, SA, and B. subtilis 10–4 + SA, both under normal and combined drought conditions, notably reduced (by 50–80% or more) the incidence of FRR development in wheat plants, with the most notable effect for B. subtilis 10–4 + SA (wherein disease symptoms were almost absent). Moreover, B. subtilis 10–4, SA, and especially B. subtilis 10–4 + SA increased plant growth (root and shoot length, fresh and dry biomass) under normal (up to 20–50%), drought (up to 15–40%), FRR (up to 15–30%), and combined drought + FRR stresses (up to 20%), with the maximum effect for B. subtilis 10–4 + SA. Additionally, B. subtilis 10–4, SA, and B. subtilis 10–4 + SA decreased stress (drought, FRR, and combined drought + FRR)-instigated lipid peroxidation and osmotic damages of plant cells. The findings indicate that endophytic bacteria B. subtilis 10–4 alone and in a mixture with SA may be used as an effective eco-friendly agent to improve wheat growth and tolerance under the influence of drought, FRR, and combinations of these stresses

Nitric Oxide (NO) Improves Wheat Growth under Dehydration Conditions by Regulating Phytohormone Levels and Induction of the Expression of the <i>TADHN</i> Dehydrin Gene

Nitric oxide (NO) is a universal signaling molecule with important regulatory functions in the plant’s life cycle and adaptation to a wide spectrum of environmental stresses including drought. The effect of pre-sowing seed treatment with the donor of NO sodium nitroprusside (SNP, 200 μM) on wheat Triticum aestivum L. plants subjected to dehydration (PEG-8000, 12%) was investigated. SNP pretreatment stimulated germination and seedling growth in normal conditions and protected them under dehydration. These effects were confirmed by percentage of seed germination, changes in fresh and dry weight of 5–6-day-old seedlings, as well as by seedlings’ linear dimensions, visual appearance, and mitotic index of the root apical meristem. Assessment of the transpiration intensity (TI) and relative water content (RWC) showed that SNP pretreatment helped to maintain the water status of seedlings subjected to dehydration stress. The data obtained by enzyme-linked immunosorbent assay (ELISA) suggested that the positive effects of SNP may be due to its influence on the phytohormonal system. SNP pretreatment induced an increase in the level of indolylacetic acid (IAA) and especially cytokinins (CK), while essential changes in ABA content were not detected. Water deficiency caused a substantial increase in ABA content and a decrease in the levels of CK and IAA. Pre-sowing SNP treatment decreased stress-induced fluctuations in the content of all studied phytohormones. Using reverse-transcription PCR (RT-PCR), we obtained data on the increase in expression of the TADHN dehydrin gene in SNP-pretreated seedlings under normal and, especially, under dehydration conditions. These findings may indicate the participation of dehydrins in NO-induced defense reactions in wheat plants under water stress. Furthermore, exogenous NO had a stabilizing effect on membrane cellular structures, as evidenced by the reduction of electrolyte leakage (EL) levels and malondialdehyde (MDA) content in dehydrated wheat seedlings under the influence of pre-sowing SNP treatment

Novel Approaches for Sustainable Horticultural Crop Production: Advances and Prospects

Reduction of plant growth, yield and quality due to diverse environmental constrains along with climate change significantly limit the sustainable production of horticultural crops. In this review, we highlight the prospective impacts that are positive challenges for the application of beneficial microbial endophytes, nanomaterials (NMs), exogenous phytohormones strigolactones (SLs) and new breeding techniques (CRISPR), as well as controlled environment horticulture (CEH) using artificial light in sustainable production of horticultural crops. The benefits of such applications are often evaluated by measuring their impact on the metabolic, morphological and biochemical parameters of a variety of cultures, which typically results in higher yields with efficient use of resources when applied in greenhouse or field conditions. Endophytic microbes that promote plant growth play a key role in the adapting of plants to habitat, thereby improving their yield and prolonging their protection from biotic and abiotic stresses. Focusing on quality control, we considered the effects of the applications of microbial endophytes, a novel class of phytohormones SLs, as well as NMs and CEH using artificial light on horticultural commodities. In addition, the genomic editing of plants using CRISPR, including its role in modulating gene expression/transcription factors in improving crop production and tolerance, was also reviewed

Effects of <i>Rhizobium leguminosarum</i> 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

Improving the Biocontrol Potential of Endophytic Bacteria <i>Bacillus subtilis</i> with Salicylic Acid against <i>Phytophthora infestans</i>-Caused Postharvest Potato Tuber Late Blight and Impact on Stored Tubers Quality

Potato (Solanum tuberosum L.) tubers are a highly important food crop in many countries due to their nutritional value and health-promoting properties. Postharvest disease caused by Phytophthora infestans leads to the significant decay of stored potatoes. The main objective of this study was to evaluate the effects of the endophytic bacteria, Bacillus subtilis (strain 10–4), or its combination with salicylic acid (SA), on some resistance and quality traits of stored Ph. infestans-infected potato tubers. The experiments were conducted using hydroponically grown potato mini-tubers, infected prior to storage with Ph. infestans, and then coated with B. subtilis, alone and in combination with SA, which were then stored for six months. The results revealed that infection with Ph. infestans significantly increased tuber late blight incidence (up to 90–100%) and oxidative and osmotic damage (i.e., malondialdehyde and proline) in tubers. These phenomena were accompanied by a decrease in starch, reducing sugars (RS), and total dry matter (TDM) contents and an increase in amylase (AMY) activity. Moreover, total glycoalkaloids (GA) (α-solanine, α-chaconine) notably increased in infected tubers, exceeding (by 1.6 times) permissible safe levels (200 mg/kg FW). Treatments with B. subtilis or its combination with SA decreased Ph. infestans-activated tuber late blight incidence (by 30–40%) and reduced oxidative and osmotic damages (i.e., malondialdehyde and proline) and AMY activity in stored, infected tubers. Additionally, these treatments decreased pathogen-activated GA accumulation and increased ascorbic acid in stored tubers. Thus, the results indicated that endophytic bacteria B. subtilis, individually, and especially in combination with SA, have the potential to increase potato postharvest resistance to late blight and improve tuber quality in long-term storage