Drought-tolerant Pseudomonas sp. showed differential expression of stress-responsive genes and induced drought tolerance in Arabidopsis thaliana.

The growth and persistence of rhizobacteria in soils are highly impacted by moisture stress. In this study, we report the first transcript analysis of four Pseudomonas strains (PS1, PS2, PS3, and PS4) isolated from the root-soil interface of rice and maize associated with different moisture levels during water deprivation. Filtered Pseudomonas sp. cells incubated at low (RH10%) and high (RH85%) relative humidity showed decreased survival of all Pseudomonas sp. at RH10% when compared with RH85%. RT-PCR showed differential expression of treS (trehalose synthase), rpoS (sigma factor), mucA (alginate regulatory gene), and fliM (flagellar motor switch protein gene) in response to exposure to RH10%. However, molecular fingerprinting and nutrient assimilation profile of Pseudomonas strains demonstrated genetic and physiological variation between the four strains irrespective of water regime and host. In vitro testing of these strains showed ACC deaminase activity and gibberellic acid, abscisic acid, indole acetic acid, and exopolysaccharide production. We determined that 50 µl of 1.2 × 103 CFU ml-1 of these Pseudomonas strains was enough to protect Arabidopsis plants against drought stress in a pot experiment. Inoculated plants increased their root colonization ability and biomass; however, PS2 showed higher survival (95%), relative water content (59%), chlorophyll (30%), glycine betaine (38%), proline (23%), and reduced MDA (43%) in shoots than irrigated control under induced water deprivation. It can be concluded that all Pseudomonas strains were effective in mitigating drought stress, however, PS2 appears to impart more resistance to drought than the other strains by upregulating key defense mechanisms.

Drought-tolerant Bacillus megaterium isolated from semi-arid conditions induces systemic tolerance of wheat under drought conditions.

KEY MESSAGE: A detailed study of the response of wheat plants, inoculated with drought-tolerant PGPR is studied which would be beneficial to achieve genetic improvement of wheat for drought tolerance. Drought stress, a major challenge under current climatic conditions, adversely affects wheat productivity. In the current study, we observed the response of wheat plants, inoculated with drought-tolerant plant growth-promoting rhizobacteria (PGPR) Bacillus megaterium (MU2) and Bacillus licheniformis (MU8) under induced drought stress. In vitro study of 90 rhizobacteria exhibited 38 isolates showed one or more plant growth-promoting properties, such as solubilization of phosphorus, potassium, and exopolysaccharide production. Four strains revealing the best activities were tested for their drought-tolerance ability by growing them on varying water potentials (- 0.05 to - 0.73 MPa). Among them, two bacterial strains Bacillus megaterium and Bacillus licheniformis showed the best drought-tolerance potential, ACC deaminase activities, IAA production, and antagonistic activities against plant pathogens. Additionally, these strains when exposed to drought stress (- 0.73 MPa) revealed the induction of three new polypeptides (18 kDa, 35 kDa, 30 kDa) in Bacillus megaterium. We determined that 106 cells/mL of Bacillus megaterium and Bacillus licheniformis were enough to induce drought tolerance in wheat under drought stress. These drought-tolerant strains increased the germination index (11-46%), promptness index (16-50%), seedling vigor index (11-151%), fresh weight (35-192%), and dry weight (58-226%) of wheat under irrigated and drought stress. Moreover, these strains efficiently colonized the wheat roots and increased plant biomass, relative water content, photosynthetic pigments, and osmolytes. Upon exposure to drought stress, Bacillus megaterium inoculated wheat plants exhibited improved tolerance by enhancing 59% relative water content, 260, 174 and 70% chlorophyll a, b and carotenoid, 136% protein content, 117% proline content and 57% decline in MDA content. Further, activities of defense-related antioxidant enzymes were also upregulated. Our results revealed that drought tolerance was more evident in Bacillus megaterium as compared to Bacillus licheniformis. These strains could be effective bioenhancer and biofertilizer for wheat cultivation in arid and semi-arid regions. However, a detailed study at the molecular level to deduce the mechanism by which these strains alleviate drought stress in wheat plants needs to be explored.

Volatile organic compounds produced by Pseudomonas pseudoalcaligenes alleviated drought stress by modulating defense system in maize (Zea mays L.).

Research on plant growth-promoting bacteria (PGPR) revealed an effective role of bacterial volatile organic compounds (VOCs) in stress alleviation. Out of 15 PGPR strains, infection with VOCs from Pseudomonas pseudoalcaligenes' resulted in maximum germination, growth promotion, and drought tolerance in maize plants. The VOCs of P. pseudoalcaligenes caused induced systemic tolerance in maize plants during 7 days of drought stress. The VOCs exposed plants displayed resistance to drought stress by reducing electrolyte leakage and malondialdehyde content and increasing the synthesis of photosynthetic pigments, proline, and phytohormones contents. Maize plants revealed enhanced resistance by showing higher activities of antioxidant defense enzymes both in shoots and roots under drought stress. Activities of antioxidant enzymes were more pronounced in shoots than roots. Gas chromatography and mass spectrophotometric (GC-MS) analysis comparing VOCs produced by the most efficient P. pseudoalcaligenes strain and inefficient strains of Pseudomonas sp. grown in culture media revealed nine compounds that they had in common. However, dimethyl disulfide, 2,3-butanediol, and 2-pentylfuran were detected only in P. pseudoalcaligenes, indicating these compounds are potential candidates for drought stress induction. Further studies are needed to unravel the molecular mechanisms of VOCs-mediated systemic drought tolerance in plants related to each identified VOC.

Combined application of zinc oxide nanoparticles and biofertilizer to induce salt resistance in safflower by regulating ion homeostasis and antioxidant defence responses.

Salinity is a key devastating abiotic factor that hinders the development and yield of safflower. The sole and combined application of zinc oxide nanoparticles (ZnO-NPs) and a biofertilizer (BF) to improve salt tolerance in safflower has not been thoroughly explored. The response of safflower plants in a pot experiment to the foliar spray of ZnO-NPs alone and in combination with a BF was thus detected. We determined that a ZnO-NP concentration of 17 mg/L was sufficient to protect safflower against salinity (250 mM NaCl) by increasing the plant productivity, percent water content, and osmolyte levels. Coapplication of ZnO-NPs and Phytoguard protected safflower plants from salinity stress by improving the activities of antioxidant enzymes and decreasing the levels of proline (leaves (61%) and roots (63%)) and malondialdehyde (MDA) (leaves (54%) and roots (65%)). Under salt stress, the Na+ content increased, while seed coating with biofertilizer and ZnO-NP spray significantly decreased the Na+ concentration (74% in leaves and 60% in roots). For the K+ concentration, however, antagonistic outcomes were observed. Additionally, the combined treatment significantly enhanced agronomic parameters such as the number of leaves and pods per plant, capitulum weight, and the number of yellow and wilted leaves per plant under salinity stress. Thus, ZnO-NPs could be an effective bio-source for the protection of safflower plants under salinity stress. Our findings showed that in the combined treatment of ZnO-NPs and biofertilizer, the salinity tolerance was more pronounced than in the single treatment and untreated control. A thorough analysis at the molecular level, however, is still required to understand the mechanism by which ZnO-NPs and BF in safflower plants alleviate salt stress.

Multitrait Pseudomonas spp. Isolated from Monocropped Wheat (Triticum aestivum) Suppress Fusarium Root and Crown Rot.

Fusarium root and crown rot is the most common disease of wheat, especially wheat grown in arid zones where drought is a common issue. The development of environmentally safe approaches to manage diseases of food crops is important for humans. The monocropping system recruits beneficial bacteria that promote plant growth through nutrient solubilization and pathogen suppression. In this study, a field where wheat was monocropped for 5 successive years under rainfed conditions was identified. A total of 29 bacterial isolates were obtained from the rhizosphere, endosphere, and phyllosphere of wheat at its harvesting stage. The Gram-negative bacteria were less prevalent (41%) but the majority (75%) exhibited plant growth-promoting traits. The ability of strains to solubilize nutrients (solubilization index = 2.3 to 4), inhibit pathogenic fungi (25 to 56%), and produce antifungal compounds was highly variable. The rhizobacteria significantly promoted the growth and disease resistance of wheat varieties such as Pirsbak-2015 and Galaxy-2013 by inducing antioxidant enzyme activity (0.2- to 2.1-fold). The bacterial strains were identified as Ochrobactrum spp., Acinetobacter spp., and Pseudomonas mediterranea by 16S rRNA and rpoD sequence analysis. The endophytic bacterium P. mediterranea HU-9 exhibited maximum biocontrol efficacy against wheat root and crown rot diseases with a disease score/disease index from 1.8 to 3.1. The monocropping systems of rainfed agriculture are an ideal source of beneficial bacteria to use as bioinoculants for different crops.

Nutritional value of Sesamum indicum L. was improved by Azospirillum and Azotobacter under low input of NP fertilizers.

BACKGROUND: Sesame (Sesame indicum L.) is well-known as a versatile industrial crop having various usages and contains 50-55% oil, 20% protein, 14-20% carbohydrate and 2-3% fiber. Several environmental factors are known to adversely affect yield and productivity of sesame. Our overall aim was to improve the growth, yield and quality of sesame cv. TS-3 using plant growth promoting rhizobacteria (PGPR) and saving the nitrogen and phosphate fertilizers (NP) by 50%. Field experiment (randomized complete block design) was conducted during the months of July to October of two consecutive years 2012-2013. Azospirillum (AL) and Azotobacter (AV) were applied as seed inoculation alone as well as along with half of the recommended dose of nitrogen (N) and phosphate (P) fertilizers (urea and diammonium phosphate) at the rate of 25 kg/ha and 30 kg/ha respectively. RESULTS: Here we report that A. lipoferum along with half dose of NP fertilizers (ALCF) were highly effective in increasing the agronomic and yield traits of sesame as compared to the control. A. vinelandii plus NP fertilizers (AVCF) exhibited higher seed oil content. Minimum acid value, optimum specific gravity and modified fatty acid composition were observed in ALCF treatment. Increase in oleic acid by ALCF is directly linked with improved oil quality for health benefits as oleic acid is the fatty acid which creates a balance between saturation and unsaturation of oil and for the hypotensive (blood pressure reducing) effects. CONCLUSION: It is inferred that ALCF treatment improved plant growth, seed yield and oil quality of sesame pertaining to good quality edible oil production.

Biocontrol of Rhizoctonia solani in basmati rice by the application of Lactobacillus and Weissella spp.

Rice is a staple food crop and is a major source of employment and income in the world. However, the attack of fungal disease poses a serious threat to the crop growth and productivity and leads toward yield loses. Therefore, current study was performed to evaluate the biocontrol potential of Lactobacillus and Weissella spp. on basmati rice against Rhizoctonia solani. Agar disc method was performed to evaluate the antifungal activity of both bacteria against R. solani. Petri plate and pot experiments were conducted to evaluate the growth promotion and biocontrol potential of both bacteria in Basmati rice under R. solani stress. Results indicated that maximum antifungal activity (82%) was recorded by Lactobacillus sp. Maximum phosphate solubilization and siderophore production was recorded by Weissella sp. In petri plate experiment, maximum root length, root fresh and dry weight (36%, 40% and 13%) was recorded by Weissella sp. and maximum shoot length and shoot fresh weight (99% and 107%) by Lactobacillus sp. In pot experiment, both bacteria enhanced the growth parameters of Basmati rice including root and shoot length, fresh and dry weight as well as no. of lateral roots. Application of Weissella sp. resulted in maximum increase (332% and 134%) in chlorophyll a and b content while Lactobacillus sp. + R. solani showed maximum (42%) carotenoid contents. Lactobacillus sp. + R. solani showed maximum increase in the proline (54%) and sugar contents (100%) while Lactobacillus sp. alone showed maximum (35%) soluble protein contents. Plant defense enzymes i-e SOD (400%), POD (25%), CAT (650%), PPO (14%) and PAL (124%) were notably increased by Weissella sp. + R. solani and Lactobacillus sp + R. solani. The Lactobacillus sp showed the best results in antifungal activity against R. solani and Weissella sp. showed the best results in production of defense enzymes in basmati rice against R. solani stress and can be suggested as the potent biocontrol agents for the rice crop.

Zinc oxide nanoparticles mediated biostimulant impact on cadmium detoxification and in silico analysis of zinc oxide-cadmium networks in Zea mays L. regulome.

Cadmium (Cd) toxicity can significantly limit plant growth and development. To eliminate the toxic effects of Cd stress, we intended to evaluate the biochemical mediated physiological responses in maize treated with biostimulant and zinc oxide nanoparticles (ZnPs). In silico analysis exhibited that the maize treated with Cd stress (200 µM) had an adverse impact on CAT1, CAT2, CAT3 and gor1 proteins, which are influential in managing the machinery of redox homeostasis. While maize inoculated with bacteria-based biostimulant and ZnPs (10 ppm) showed prominently improved biomass, chlorophyll a, b and carotenoid content. We found a significant increase in the total sugar, protein, proline content and antioxidants under the effect of Cd stress. However, these parameters are further enhanced by applying biostimulants and ZnPs. Declined lipid peroxidation and membrane solubilization index under the effect of biostimulant and ZnPs was observed. Furthermore, these treatments improved maize's zinc, copper, sodium, magnesium, iron, potassium and calcium content. Based on these results, an antagonistic relationship between Zn and Cd uptake that triggered efficient Cd detoxification in maize shoot was found. Scanning electron micrography showed distorted leaf structure of the Cd stressed plants while the biostimulant and ZnPs reduced the structural cell damage of maize leaves. In silico study showed that ZnO positively regulates all protein interactors, including GRMZM2G317386_P01 (Metallo endo proteinase 1-MMP), GRMZM2G110220_P01 (Metallo endo proteinase 5-MMP), GRMZM2G103055_P01 (Alpha-amylase) and GRMZM2G006069_P01 (Zn-dependent exo peptidase superfamily) proteins which are involved in energy generating processes, channels formation, matrix re-localization and stress response. This suggests that ZnO offers an ideal role with protein interactors in maize. Our findings depict that these treatments, i.e., biostimulant and ZnPs alone, are efficient enough to exhibit Cd remediation potential in maize; however, their combination showed synergistic effects.

Coactive role of zinc oxide nanoparticles and plant growth promoting rhizobacteria for mitigation of synchronized effects of heat and drought stress in wheat plants.

This study intended to investigate the potential of the plant growth-promoting rhizobacteria (PGPR) and green synthesized zinc oxide nanoparticles (ZnO-NPs) (fruit extract of Papaya) against heat and drought stress in wheat. The characterization of green-synthesized ZnO-NPs was done through UV-vis spectrophotometry, Fourier-transform infrared spectrometry, X-ray diffraction and scanning electron microscopy. Individual and combination of PGPR (Pseudomonas sp.) and ZnO-NPs (10 ppm) amendments were tested in a pot experiment to upregulate wheat defence system under three stress groups (drought, heat and combined heat and drought stress). Drought and heat stress synergistically caused higher damage to wheat plants than individual heat and drought stress. This observation was confirmed with remarkable higher MDA and hydrogen peroxide (H2O2) content. Treated plants exposed to all stress groups showed an improved wheat growth and stress resistance through better biomass, photosynthetic pigments, nutrients, soluble sugars, protein and indole acetic acid content. Combination of ZnO-NPs and Pseudomonas sp. Protects the plants from all stress groups by producing higher proline, antioxidant enzymes i. e superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, glutathione reductase and dehydroascorbate reductase, and abscisic acid. Moreover, higher stress alleviation by this treatment was manifested by marked reduced electrolyte leakage, MDA and H2O2. The findings of current study confirmed that the synergistic actions of PGPR and ZnO-NPs can rescue plants from both single and combined heat and drought stress.

Heavy Metal-Resistant Plant Growth-Promoting Citrobacter werkmanii Strain WWN1 and Enterobacter cloacae Strain JWM6 Enhance Wheat (Triticum aestivum L.) Growth by Modulating Physiological Attributes and Some Key Antioxidants Under Multi-Metal Stress.

Due to wastewater irrigation, heavy metal (HM) exposure of agricultural soils is a major limiting factor for crop productivity. Plant growth-promoting bacteria (PGPB) may lower the risk of HM toxicity and increase crop yield. In this context, we evaluated two HM-resistant PGPB strains, i.e., Citrobacter werkmanii strain WWN1 and Enterobacter cloacae strain JWM6 isolated from wastewater-irrigated agricultural soils, for their efficacy to mitigate HM (Cd, Ni, and Pb) stress in a pot experiment. Increasing concentrations (0, 50, 100, and 200 ppm) of each HM were used to challenge wheat plants. Heavy metal stress negatively affected wheat growth, biomass, and physiology. The plants under elevated HM concentration accumulated significantly higher amounts of heavy metals (HMs) in shoots and roots, resulting in increased oxidative stress, which was evident from increased malondialdehyde (MDA) content in roots and shoots. Moreover, alterations in antioxidants like superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT) were observed in plants under HM stress. The severity of damage was more pronounced with rising HM concentration. However, inoculating wheat with Citrobacter werkmanii strain WWN1 and Enterobacter cloacae strain JWM6 (107 CFU ml-1) improved plant shoot length (11-42%), root length (19-125%), fresh weight (41-143%), dry weight (65-179%), and chlorophyll a (14%-24%) and chlorophyll b content (2-24%) under HM stress. Citrobacter werkmanii strain WWN1 and Enterobacter cloacae strain JWM6 either alone or in co-inoculation enhanced the antioxidant enzyme activity, which may lower oxidative stress in plants. However, seeds treated with the bacterial consortium showed an overall better outcome in altering oxidative stress and decreasing HM accumulation in wheat shoot and root tissues. Fourier transform infrared spectroscopy indicated the changes induced by HMs in functional groups on the biomass surface that display effective removal of HMs from aqueous medium using PGPB. Thus, the studied bacterial strains may have adequate fertilization and remediation potential for wheat cultivated in wastewater-irrigated soils. However, molecular investigation of mechanisms adopted by these bacteria to alleviate HM stress in wheat is required to be conducted.

Comparative analysis of iron oxide nanoparticles synthesized from ginger (Zingiber officinale) and cumin seeds (Cuminum cyminum) to induce resistance in wheat against drought stress.

In the present study, iron oxide nanoparticles (Fe3O2-NPs) synthesized from ginger (Zingiber officinale) and cumin seeds (Cuminum Cyminum L.) extracts were investigated to reveal their potential to enhance the growth and drought resistance of wheat plants under drought stress. In an In Vitro experiment, four different concentrations for Fe3O2-NPs (0.3 mM, 0.6 mM, 0.9 mM, and 1.2 mM) of ginger and cumin seeds were tested. Among all the concentrations tested, ginger Fe3O2-NPs (0.6 mM) and cumin seeds Fe3O2-NPs (1.2 mM) were more effective to enhance wheat germination, biomass, and survival percentage under drought stress and irrigated conditions than the non-treated control plant. In a pot experiment, wheat plants under induced water stress showed marked up-regulation in the biochemical resistance mechanisms when treated with ginger Fe3O2-NPs (0.6 mM) and cumin seeds Fe3O2-NPs (1.2 mM) than the non-treated control. Cumin seeds Fe3O2-NPs (1.2 mM) were more effective than ginger Fe3O2-NPs (0.6 mM) in ameliorating adverse effects of drought stress in wheat. Results demonstrated that cumin seeds Fe3O2-NPs (1.2 mM) exhibited a higher increase in chlorophyll a, b and carotenoids (72%, 265% and 96% respectively), proline (127%), superoxide dismutase (115%), peroxidase (43.8%), ascorbate peroxidase (44.6%). This also showed higher reduction in lipid peroxidation, electrolyte leakage and increased soluble sugars and total Fe content in the roots and shoots than non-treated plants under drought. Hence, nano-priming can be considered an effective strategy for sustainable food production in marginal soils.

Phosphate solubilizing bacteria enhanced growth, oil yield, antioxidant properties and biodiesel quality of Kasumbha.

Biodiesel is considered as a potential alternative energy source, but problem exists with the quantity and quality of feedstock used for it. To improve the feedstock quality of biodiesel, a field experiment was conducted under natural conditions. Cultivar Thori of kasumbha was used in the experiment. Commercialized biofertilizers were applied at the rate of 20 kg per acre and chemical fertilizer (diammonium phosphate) was applied as half dose (15 kg/ha). Results indicated that number of leaf plant-1, leaf area, number of seeds capitulum-1 was significantly increased by biofertilizer treatment alone (BF) and combine treatment of biofertilizer and chemical fertilizer (BFCF). Agronomic traits such as plant height, no. of branches of a plant, no. of capitulum/plant was improved significantly by BF treatment over the control. Maximum 1000 seed weight (41%) and seed yield (23%) were recorded in half dose of chemical fertilizers treatment (CFH). Seed oil content and seed phenolics were significantly improved by BF and CF treatments while maximum biodiesel yield was recorded by BF treatment. Maximum oleic acid was recorded by BF treatment while other fatty acids being maximum in control except linoleic acid in BFCF treatment. Results for specific gravity were non-significant while acid value and free fatty acid contents were substantially reduced by BF treatment as compared to other treatments. Maximum value of iodine number was recorded in BFCF treatment while tocopherol contents were improved by BF treatment. It is inferred that biofertilizer treatment alone perform better as compared to other treatments and 50% chemical fertilizer can be replaced using biofertilizer which is a good approach for sustainable environmental-friendly agriculture.

Prevalence of Antimicrobial Resistance and Respective Genes among Bacillus spp., a Versatile Bio-Fungicide.

The plant rhizosphere is not only a reservoir of microbes but also a hub of antimicrobial resistance genes. Rhizospheric Bacillus spp. are the potential bio-inoculants with a versatile application in agriculture as bio-fertilizer and bio-fungicide. In the current study, the potential bio-control agent that is the Bacillus species (n = 7) was screened for the antimicrobial resistance pattern to assess their risk before registering them as a bio-inoculant. All of the Bacillus spp. were categorized as multi-drug-resistant (MDR), bacteria but none of them was either pan-drug-resistant (PDR) or extensive-drug-resistant (XDR). The multiple antimicrobial resistance (MAR) index of Bacillus spp. was higher than the critical value (0.2). The Bacillus spp. showed resistance to antimicrobial classes such as ß lactam, macrolides, sulfonamides, tetracycline, aminoglycosides, and lincosamide. Various antimicrobial resistance genes, namely VmiR, ImrB, tetL, mphK, ant-6, penp, and bla OXA, associated with different mechanisms of resistance, were also detected in Bacillus spp. The Bacillus spp. also showed stress-tolerance traits such as ACC deaminase and EPS activity except the strains MAZ-117 and FZV-34, respectively. A significant correlation was observed between the PGPR and antimicrobial resistance, which shows that they may have adapted drug-resistance mechanisms to tolerate the environmental stress. These findings suggest that bio-fungicidal Bacillus spp. could be used very carefully on a commercial scale.

Alleviation of cadmium stress in rice by inoculation of Bacillus cereus.

Heavy metal resistant bacteria are of great importance because they play a crucial role in bioremediation. In the present study, 11 bacterial strains isolated from industrial waste were screened under different concentrations of cadmium (Cd) (100 µM and 200 µM). Among 11 strains, the Cd tolerant Bacillus cereus (S6D1-105) strain was selected for in vitro and in vivo studies. B. cereus was able to solubilize potassium, and phosphate as well as produce protease and siderophores during plate essays. Moreover, we observed the response of hydroponically grown rice plants, inoculated with B. cereus which was able to promote plant growth, by increasing plant biomass, chlorophyll contents, relative water content, different antioxidant enzymatic activity such as catalase, superoxide dismutase, ascorbate peroxidase, polyphenol oxidase and phenylalanine ammonia-lyase and reducing malondialdehyde content in both roots and leaves of rice plants under Cd stress. Our results showed that the B. cereus can be used as a biofertilizer which might be beneficial for rice cultivation in Cd contaminated soils.

Pseudomonas spp. Mediate defense response in sugarcane through differential exudation of root phenolics.

Pseudomonas spp., a ubiquitous biocontrol agent, protects the plants from phytopathogens by suppressing them directly by reinforcing the plant's intrinsic defense mechanism. Root exudated phenolics play an important role in establishing the rhizobacteria population and cross the host boundaries in beneficial plant-microbe interaction. In this study, Pseudomonas spp. HU-8 & HU-9 antagonized the sugarcane red rot pathogen (C. falcatum) and showed a positive chemotactic response against different concentrations (10-30 µM) of synthetic phenolic acids like p-coumaric, vanillic, and 3,4 di-hydroxybenzoic acid. In a pot experiment, they effectively colonized the sugarcane rhizosphere and mediated defense response in sugarcane plants challenged with red rot pathogen C. falcatum by regulating the exudation of root phenolics under hydroponic conditions. They significantly induced the activity of the antioxidant enzymes CAT (1.24-1.64 fold), PO (0.78-1.61 fold), PAL (0.77-0.97 fold), and PPO (3.67-3.73 fold) over untreated plants in sugarcane. They also induced the total phenolic contents (TPC) in sugarcane in the presence (6.56-10.29 mg/g GAE) and absence (2.89-4.16 mg/g GAE) of the pathogen quantified through the Folin-Ciocalteu (FC) method. However, their effect was lower than that of the pathogen (4.34-8 mg/g GAE). The Pseudomonas spp. significantly colonized the sugarcane rhizosphere by maintaining a cell population of (1.0E + 07-1.3E + 08 CFU/mL). A significant positive Pearson's correlation was observed between the root exudated total phenolic contents, antioxidant enzymatic activities, and rhizospheric population of inoculated bacteria. The 16S rRNA and rpoD gene analysis showed sequence conservation (C: 0.707), average number of nucleotide differences (k: 199.816), nucleotide diversity, (Pi): 0.09819), average number of informative nucleotide sites per site (Psi: 0.01275), GC content (0.57), and polymorphic sites (n = 656). These diverse Pseudomonas spp. could be an ideal bio-inoculants for a broad range of hosts especially graminaceous crops.

Exogenous silicon and hydrogen sulfide alleviates the simultaneously occurring drought stress and leaf rust infection in wheat.

Silicon (Si) and hydrogen sulfide (H2S) are known to enhance plant defense against multiple stresses. Current study was conducted to investigate the application of Si and H2S alone as well as in combination, improved physiological resilience of wheat plants to drought stress (DS) and pathogen-Puccinia triticina (Pt) infection. We aimed to increase the wheat plant growth and to enhance the DS tolerance and Pt resistance with the concurrent applications of H2S and Si. In the first experiment, we selected the best growth enhancing concentration of H2S (0.3 mM) and Si (6 mM) to further investigate their tolerance and resistance potential in the pot experiment under DS and pathogen infection conditions. The obtained results reveal that DS has further increased the susceptibility of wheat plants to leaf rust pathogen infection while, the sole application of Si and the simultaneous exogenous treatments of H2S + Si enhanced the plant growth, decreased disease incidence, and significantly improved tolerance and defense mechanisms of wheat under individual and interactive stress conditions. The exogenous treatment of H2S + Si improved the growth criteria, photosynthetic pigments, osmoprotectants, and defense related enzyme activities. The same treatment also reinforced the endogenous H2S, Si, ABA and SA contents while decreased the disease incidence and oxidative stress indicators under individual and combined stress conditions. Overall, results from this study presents the influence of combined drought and P. triticina stress in wheat and reveal the beneficial impacts of concurrent exogenous treatment of H2S + Si to mitigate the drought and pathogen (P. triticina) induced adverse effects.

Prevalence of Wheat Associated Bacillus spp. and Their Bio-Control Efficacy Against Fusarium Root Rot.

Bacillus spp. are the most prevalent group of bacteria in nature. Their prevalence depends upon multiple factors, namely, sporulation, antagonism, and production of secondary metabolites. The development of an eco-friendly approach to cope with edible crops diseases is very substantial for humans. In the present study, 658 isolates were obtained from wheat grown in the wheat rice cropping system and tested for their antagonistic activity against four wheat root rot pathogens, namely, Fusarium oxysporum, Fusarium moniliforme, Macrophomina phaseolina, and Rhizoctonia solani. Out of 658, 106 isolates were found antagonistic to either single or multiple fungi. Out of 106 antagonistic bacteria, 62 (23%) were rhizospheric, 28 (14%) were root endospheric, and 16 (9%) were leaf endospheric. Based on mean inhibition against all fungi, the bacterial strains SM-39 and SM-93 showed maximum antagonistic activity. The 16S rRNA gene analysis revealed that most of the antagonistic bacteria exhibiting ≥48% antagonism were Bacillus spp. (98%), except two were Klebsiella spp. (2%). The bacterial strains exhibited phylogenetic lineage with the type strains of the respective genus based on the 16S rRNA gene sequences. In the net house experiment, Bacillus velezensis (SM-39) and Bacillus cabrialesii (SM-93) significantly suppressed Fusarium root rot severity in wheat (42-62%). Plants treated with these strains had lower electrolytic leakage (29-36%), as compared to untreated (44%). Relative water content was much higher (46-58%) for plants inoculated with these strains. These antagonistic strains also considerably colonized the wheat rhizosphere with a cell population of 5.8-6.9.log CFU/g of soil. The rhizosphere of wheat grown in the wheat-rice cropping system could be the potential habitat of effective biocontrol agents.

Antagonistic, Anti-oxidant, Anti-inflammatory and Anti-diabetic Probiotic Potential of Lactobacillus agilis Isolated From the Rhizosphere of the Medicinal Plants.

Potential probiotic bacteria can be used as a biotherapeutic agent and a sustainable alternative to antibiotics, as an anti-oxidative, anti-inflammatory, and anti-diabetic agent without causing any serious side effects. Mostly human-friendly Lactic acid bacteria (LAB) have been isolated from the animal-human origin to be used as biotherapeutic agents or to produce useful metabolites (nutraceutical). However, less information is known about the role of medicinal plants associated LAB as biotherapeutic agents. The isolation of 115 human-friendly Lactobacillus strains was done from the rhizosphere of the medicinal plants Ocimum tenuiflorum, Azadirachta indica, Ficus carica. The obtained bacteria were then tested for their safe status before being using it for a beneficial purpose. Out of 115 strains, 29 (25%) were negative for blood hemolytic activities. Among these 29 isolates, three isolates did not show a breakdown of gelatin and were recognized as safe. Antibiotic resistance assay showed resistance of two of them against antibiotics discs of Streptomycin (10 µg), Ciprofloxacin (20 µg), Vancomycin (30 µg), Metronidazole (10 µg), Ampicillin (5 µg), Chloramphenicol (30 µg), Kanamycin (30 µg), Erythromycin (15 µg), Penicillin (10 µg) and Tetracycline (30 µg). The bacterial isolate (T-2) was found safe that was identified as Lactobacillus agilis by sequence analysis of 16 s rRNA gene and processed in vitro as an anti-bacterial, anti-oxidant, anti-diabetic, and anti-inflammatory agent. Free radical scavenging activities and inhibition of α-amylase activities for Lactobacillus agilis were found relative to standard drug values as 68% and 73% and 51.3% and 65.3%, respectively. The in-vitro anti-inflammatory assay showed 61.6% (Lactobacillus agilis) while showed 69% (aspirin) activity for denaturation albumin protein. The results suggested that Lactobacillus agilis can be used as a potential probiotic strain as well as can be used to produce nutraceuticals.

Sodium nitroprusside application improves morphological and physiological attributes of soybean (Glycine max L.) under salinity stress.

Salinity is among the major abiotic stresses negatively affecting the growth and productivity of crop plants. Sodium nitroprusside (SNP) -an external nitric oxide (NO) donor- has been found effective to impart salinity tolerance to plants. Soybean (Glycine max L.) is widely cultivated around the world; however, salinity stress hampers its growth and productivity. Therefore, the current study evaluated the role of SNP in improving morphological, physiological and biochemical attributes of soybean under salinity stress. Data relating to biomass, chlorophyll and malondialdehyde (MDA) contents, activities of various antioxidant enzymes, ion content and ultrastructural analysis were collected. The SNP application ameliorated the negative effects of salinity stress to significant extent by regulating antioxidant mechanism. Root and shoot length, fresh and dry weight, chlorophyll contents, activities of various antioxidant enzymes, i.e., catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) were improved with SNP application under salinity stress compared to control treatment. Similarly, plants treated with SNP observed less damage to cell organelles of roots and leaves under salinity stress. The results revealed pivotal functions of SNP in salinity tolerance of soybean, including cell wall repair, sequestration of sodium ion in the vacuole and maintenance of normal chloroplasts with no swelling of thylakoids. Minor distortions of cell membrane and large number of starch grains indicates an increase in the photosynthetic activity. Therefore, SNP can be used as a regulator to improve the salinity tolerance of soybean in salt affected soils.

Halotolerant rhizobacteria Pseudomonas pseudoalcaligenes and Bacillus subtilis mediate systemic tolerance in hydroponically grown soybean (Glycine max L.) against salinity stress.

Salt stress is one of the devastating factors that hampers growth and productivity of soybean. Use of Pseudomonas pseudoalcaligenes to improve salt tolerance in soybean has not been thoroughly explored yet. Therefore, we observed the response of hydroponically grown soybean plants, inoculated with halotolerant P. pseudoalcaligenes (SRM-16) and Bacillus subtilis (SRM-3) under salt stress. In vitro testing of 44 bacterial isolates revealed that four isolates showed high salt tolerance. Among them, B. subtilis and P. pseudoalcaligenes showed ACC deaminase activity, siderophore and indole acetic acid (IAA) production and were selected for the current study. We determined that 106 cells/mL of B. subtilis and P. pseudoalcaligenes was sufficient to induce tolerance in soybean against salinity stress (100 mM NaCl) in hydroponics by enhancing plant biomass, relative water content and osmolytes. Upon exposure of salinity stress, P. pseudoalcaligenes inoculated soybean plants showed tolerance by the increased activities of defense related system such as ion transport, antioxidant enzymes, proline and MDA content in shoots and roots. The Na+ concentration in the soybean plants was increased in the salt stress; while, bacterial priming significantly reduced the Na+ concentration in the salt stressed soybean plants. However, the antagonistic results were observed for K+ concentration. Additionally, soybean primed with P. pseudoalcaligenes and exposed to 100 mM NaCl showed a new protein band of 28 kDa suggesting that P. pseudoalcaligenes effectively reduced salt stress. Our results showed that salinity tolerance was more pronounced in P. pseudoalcaligenes as compared to B. subtilis. However, a detailed study at molecular level to interpret the mechanism by which P. pseudoalcaligenes alleviates salt stress in soybean plants need to be explored.