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 strain-specifically improves growth of Phaseolus vulgaris plants under normal and salinity conditions and exerts anti-stress effect through induced lignin deposition in roots and decreased oxidative and osmotic damages.

Bacillus subtilis is one of the non-pathogenic beneficial bacteria that promote plant growth and stress tolerance. In the present study, we revealed that seed priming with endophytic B. subtilis (strains 10-4, 26D) improved Phaseolus vulgaris L. (common bean) seed germination and plant growth under both saline and non-saline conditions. 10-4 and 26D decreased oxidative and osmotic damage to the plant cells since bacterial inoculations reduced lipid peroxidation and proline accumulation in plants under salinity. 26D and especially 10-4 preserved different elevated levels of chlorophyll (Chl) a and Chl b in bean leaves under salinity, while carotenoids (Car) increased only by 10-4 and slightly decreased by 26D. Under normal conditions, 10-4 and 26D did not affect Chl a and Car concentrations, while Chl b decreased in the same plants. Under non-saline and especially saline conditions, 10-4 and 26D significantly increased lignin accumulation in plant roots and the highest lignin content along with better growth and oxidative damages reduction was observed after 10-4 inoculation under salinity, indicating a major role of B. subtilis-induced strengthening the root cell walls in the implementation protective effect of studied bacteria on plants. Therefore, B. subtilis 10-4 and 26D exerts protective effects on the growth of common bean plants under salinity by regulating plant defense mechanisms and the major role in tolerance development may contribute through the activation by B. subtilis lignin deposition in roots. The obtained data also indicates a strain-dependent efficiency of endophytic B. subtilis since strains 10-4 and 26D differently improved growth attributes and modulates cellular response reactions of the same common bean plants both under normal and salinity conditions, that generates interest for further investigations in this direction.

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.

The Effect of Endophytic Bacteria Bacillus subtilis and Salicylic Acid on Some Resistance and Quality Traits of Stored Solanum tuberosum L. Tubers Infected with Fusarium Dry Rot.

The effect of endophytic Bacillus subtilis (strains 10-4, 26D) and their compositions withsalicylic acid (SA) on some resistance and quality traits of stored potatoes infected with Fusariumdry rot were studied. The experiments were carried out on hydroponically grown Solanumtuberosum L. tubers that were infected before storage with Fusarium oxysporum and coated with B.subtilis 10-4, 26D with and without exogenous SA, and then stored for six months. It has been shownthat 10-4, 26D, 10-4 + SA, and 26D + SA reduced in different levels (up to 30-50%) the incidence ofF. oxysporum-caused dry rot (with the highest effect for 10-4 + SA). SA notably enhanced the positiveeffect of 10-4, while for 26D, such an effect was not observed. All of the tested treatments increasedamylase (AMY) and AMY inhibitors activity in infected tubers, while decreased Fusarium-inducedprotease activity (except in the case of 10-4 + SA, which promoted a slight increase) was revealed.10-4, 26D, and their compositions with SA decreased (in different degrees) the pathogen-causedlipid peroxidation, proline, and reducing sugars accumulation in potatoes after long-term storage.It was also discovered 10-4 and 26D, regardless of SA presence, decrease pathogen-inducedglycoalkaloids α-Solanine and α-Chaconine accumulation and preserved increased levels of starchand total dry matter in infected stored potatoes. The findings indicate endophytic B. subtilis and itscompositions with SA is a promising eco-friendly and bio-safe approach to cope with postharvestdecays of potato during long-term storage; however, when developing preparations-compositionsit should take into account the strain-dependent manner of B. subtilis action together with SA.

Effects of Endophytic Bacillus Subtilis and Salicylic Acid on Postharvest Diseases (Phytophthora infestans, Fusarium oxysporum) Development in Stored Potato Tubers.

Postharvest diseases of potato lead to significant food and economic losses worldwide. The exogenous application of eco-friendly methods plays an important role in the control of postharvest decay. In this work the effects of endophytic bacteria B. subtilis (10-4, 26D) were studied in the context of two application parameters: concentration, with a range between 103-108 CFU/mL tested, and synergistic effects of the signal molecule salicylic acid (SA) (0.05 mM) on potato tubers' resistance to Phytophthora infestans and Fusarium oxysporum during storage. The experiments were carried out on hydroponically grown potato (Solanum tuberosum L.) mini-tubers. This study demonstrates the suppressive effect of B. subtilis (10-4, 26D) on diseases of potato during storage and reveals that this effect happens in a dose-dependent manner, both individually and in combination with SA. The most effective concentrations of B. subtilis for suppression of both Ph. infestans and F. oxysporum are 108 CFU/mL (10-4 and 26D), 107 CFU/mL (10-4 + SA) and 106 CFU/mL (26D + SA). The ability of B. subtilis (10-4, 26D) to effectively penetrate and colonize the internal tubers' tissues when applied immediately prior to storage, and the ability of SA to accelerate these processes, have been proven. B. subtilis (10-4, 26D), individually and in compositions with SA, increased ascorbic acid content and decreased pathogen-induced proline accumulation and lipid peroxidation in tubers. This indicates a protective effect conferred to cells against reactive oxygen and an extension of aging processes, manifested by a prolonged shelf life and extended preservation of fresh appearance.