Pesticide-tolerant microbial consortia: Potential candidates for remediation/clean-up of pesticide-contaminated agricultural soil.

Reclamation of pesticide-polluted lands has long been a difficult endeavour. The use of synthetic pesticides could not be restricted due to rising agricultural demand. Pesticide toxicity has become a pressing agronomic problem due to its adverse impact on agroecosystems, agricultural output, and consequently food security and safety. Among different techniques used for the reclamation of pesticide-polluted sites, microbial bioremediation is an eco-friendly approach, which focuses on the application of resilient plant growth promoting rhizobacteria (PGPR) that may transform or degrade chemical pesticides to innocuous forms. Such pesticide-resilient PGPR has demonstrated favourable effects on soil-plant systems, even in pesticide-contaminated environments, by degrading pesticides, providing macro-and micronutrients, and secreting active but variable secondary metabolites like-phytohormones, siderophores, ACC deaminase, etc. This review critically aims to advance mechanistic understanding related to the reduction of phytotoxicity of pesticides via the use of microbe-mediated remediation techniques leading to crop optimization in pesticide-stressed soils. The literature surveyed and data presented herein are extremely useful, offering agronomists-and crop protectionists microbes-assisted remedial strategies for affordably enhancing crop productivity in pesticide-stressed soils.

Evaluation of Osmotolerant Potential of Halomonas sulfidaeris MV-19 Isolated from a Mud Volcano.

Salinity is one of the major challenges for cultivation of crops in a sustainable way because it severely affects plant growth and yield. Keeping this challenge in view, in the current study, a salt-tolerant Halomonas MV-19 was isolated from an extreme niche of mud volcano of Andaman Nicobar Island, India and identified on the basis of standard morphological, biochemical, and physiological tests and identified as Halomonas sulfidaeris strain MV-19 by 16S rRNA gene sequencing. The bacterium can grow on nutrient agar and nutrient broth supplemented with 3.5 M (≥ 20%) sodium chloride (NaCl). Sugar utilization assay revealed that H. sulfidaeris MV-19 utilizes only three sugars (dextrose, fructose, and mannose) from among twenty four tested sugars. The best growth of H. sulfidaeris MV-19 was observed in nutrient broth supplemented with 8% NaCl. When the broth was supplemented with dextrose, fructose, and mannose, the H. sulfidaeris MV-19 grew maximally in nutrient broth supplemented with 8% NaCl and 5% fructose. This strain produced exopolysaccharides (EPS) in nutrient broth supplemented with 8% NaCl and sugars (dextrose, fructose, and mannose). The EPS production was increased by 350% (three and half time) after addition of 5% fructose in nutrient broth compare with the EPS production in nutrient broth without supplemented with sugars. H. sulfidaeris MV-19 strain can produce EPS, which can help aggregate soil particle and reduced osmotic potential in soil, thus, be useful in alleviation of salinity stress in different crops cultivated in saline soils. The findings of the current investigation are expected to contribute towards effective abiotic stress management.

Fractal characterizations of MeV ion treated CaF2 thin films.

We present the morphological evolution and fractal characterizations of CaF2 thin-film surfaces modified by bombardment with 100 MeV Au+8 ions at various fluences. Atomic force microscopy (AFM) combined with line profile and two-dimensional power spectral density (2D-PSD) analysis was utilized to investigate the evolution of surface morphology as a function of fluence. The AFM images were utilized to investigate the relationship between fractal dimension, roughness exponent, lateral correlation length, and ion fluence. The surface erosion owing to sputtering was depicted using Rutherford backscattering spectrometry. The structural characteristics' dependency on fluence was explored with the help of glancing angle x-ray diffraction measurements on virgin and irradiated samples. Tensile stress calculated using a peak shift in the glancing angle x-ray diffractogram showed an increase in tensile stress with fluence that caused the surface to crack after the fracture strength of the surface was crossed. 2D-PSD analysis signified the role of sputtering over surface diffusion for the observed surface modifications. Fractal dimensions first increased and then decreased with ion fluence. The lateral correlation length decreased, while the roughness exponent increased with fluence after the threshold value.

Delineation of molecular interactions of plant growth promoting bacteria induced ß-1,3-glucanases and guanosine triphosphate ligand for antifungal response in rice: a molecular dynamics approach.

BACKGROUND: The plant growth is influenced by multiple interactions with biotic (microbial) and abiotic components in their surroundings. These microbial interactions have both positive and negative effects on plant. Plant growth promoting bacterial (PGPR) interaction could result in positive growth under normal as well as in stress conditions. METHODS: Here, we have screened two PGPR's and determined their potential in induction of specific gene in host plant to overcome the adverse effect of biotic stress caused by Magnaporthe grisea, a fungal pathogen that cause blast in rice. We demonstrated the glucanase protein mode of action by performing comparative modeling and molecular docking of guanosine triphosphate (GTP) ligand with the protein. Besides, molecular dynamic simulations have been performed to understand the behavior of the glucanase-GTP complex. RESULTS: The results clearly showed that selected PGPR was better able to induce modification in host plant at morphological, biochemical, physiological and molecular level by activating the expression of ß-1,3-glucanases gene in infected host plant. The docking results indicated that Tyr75, Arg256, Gly258, and Ser223 of glucanase formed four crucial hydrogen bonds with the GTP, while, only Val220 found to form hydrophobic contact with ligand. CONCLUSIONS: The PGPR able to induce ß-1,3-glucanases gene in host plant upon pathogenic interaction and ß-1,3-glucanases form complex with GTP by hydrophilic interaction for induction of defense cascade for acquiring resistance against Magnaporthe grisea.

Biocomputational Assessment of Natural Compounds as a Potent Inhibitor to Quorum Sensors in Ralstonia solanacearum.

Ralstonia solanacearum is among the most damaging bacterial phytopathogens with a wide number of hosts and a broad geographic distribution worldwide. The pathway of phenotype conversion (Phc) is operated by quorum-sensing signals and modulated through the (R)-methyl 3-hydroxypalmitate (3-OH PAME) in R. solanacearum. However, the molecular structures of the Phc pathway components are not yet established, and the structural consequences of 3-OH PAME on quorum sensing are not well studied. In this study, 3D structures of quorum-sensing proteins of the Phc pathway (PhcA and PhcR) were computationally modeled, followed by the virtual screening of the natural compounds library against the predicted active site residues of PhcA and PhcR proteins that could be employed in limiting signaling through 3-OH PAME. Two of the best scoring common ligands ZINC000014762512 and ZINC000011865192 for PhcA and PhcR were further analyzed utilizing orbital energies such as HOMO and LUMO, followed by molecular dynamics simulations of the complexes for 100 ns to determine the ligands binding stability. The findings indicate that ZINC000014762512 and ZINC000011865192 may be capable of inhibiting both PhcA and PhcR. We believe that, after further validation, these compounds may have the potential to disrupt bacterial quorum sensing and thus control this devastating phytopathogenic bacterial pathogen.

SHI induced surface re-organization of non-amorphisable nanodimensional fluoride thin films.

Surface re-organization in nanodimensional fluoride (LiF and BaF2) thin films is observed under dense electronic excitation produced by swift heavy ion (SHI) irradiation. The irradiation was performed at an angle of less than 15° with respect to the film surface while keeping the sample at liquid nitrogen temperature. The surface of the irradiated samples was characterized by atomic force microscopy (AFM) and scanning electron microscopy (SEM) complemented by energy dispersive X-ray spectroscopy (EDX). Detailed analyses indicate that the surface starts cracking at lower fluence. With an increase in the ion fluence, the materials shrinking and surface re-structuring lead to lamellae periodic structures. The average width of the wall decreases, while the separation and the height of the structures increase with the fluence. The composition of the lamellae walls and the gap in between were analyzed by EDX. At the highest fluence of irradiation, a strong signal of the substrate and negligible signals of F and Ba are observed between the walls of the lamellae structures, which shows that the entire deposited material is removed and the Si substrate is completely exposed to the ion beam. It is also observed that the substrate remains unaffected by SHI irradiation and does not undergo any structural transformation as evident by cross-sectional SEM micrographs. Such surface re-organization is not expected in fluoride thin films due to their non-amorphizable nature even at very high fluence SHI irradiation. The concept of grain rotation under SHI irradiation is used to explain the re-organization phenomena in such non-amorphizable materials.

Enhancing spinach (Spinacia oleracea L.) resilience in pesticide-contaminated soil: Role of pesticide-tolerant Ciceribacter azotifigens and Serratia marcescens in root architecture, leaf gas exchange attributes and antioxidant response restoration.

This study unveils the detoxification potential of insecticide-tolerant plant beneficial bacteria (PBB), i.e., Ciceribacter azotifigens SF1 and Serratia marcescens SRB1, in spinach treated with fipronil (FIP), profenofos (PF) and chlorantraniliprole (CLP) insecticides. Increasing insecticide doses (25-400 µg kg-1 soil) significantly curtailed germination attributes and growth of spinach cultivated at both bench-scale and in greenhouse experiments. Profenofos at 400 µg kg-1 exhibited maximum inhibitory effects and reduced germination by 55%; root and shoot length by 78% and 81%, respectively; dry matter accumulation in roots and shoots by 79% and 62%, respectively; leaf number by 87% and leaf area by 56%. Insecticide application caused morphological distortion in root tips/surfaces, increased levels of oxidative stress, and cell death in spinach. Application of insecticide-tolerant SF1 and SRB1 strains relieved insecticide pressure resulting in overall improvement in growth and physiology of spinach grown under insecticide stress. Ciceribacter azotifigens improved germination rate (10%); root biomass (53%); shoot biomass (25%); leaf area (10%); Chl-a (45%), Chl-b (36%) and carotenoid (48%) contents of spinach at 25 µg CLP kg-1 soil. PBB inoculation reinvigorated the stressed spinach and modulated the synthesis of phytochemicals, proline, malondialdehyde (MDA), superoxide anions (O2•-), and hydrogen peroxide (H2O2). Scanning electron microscopy (SEM) revealed recovery in root tip morphology and stomatal openings on abaxial leaf surfaces of PBB-inoculated spinach grown with insecticides. Ciceribacter azotifigens inoculation significantly increased intrinsic water use efficiency, transpiration rate, vapor pressure deficit, intracellular CO2 concentration, photosynthetic rate, and stomatal conductance in spinach exposed to 25 µg FIP kg-1. Also, C. azotifigens and S. marcescens modulated the antioxidant defense systems of insecticide-treated spinach. Bacterial strains were strongly colonized to root surfaces of insecticide-stressed spinach seedlings as revealed under SEM. The identification of insecticide-tolerant PBBs such as C. azotifigens and S. marcescens hold the potential for alleviating abiotic stress to spinach, thereby fostering enhanced and safe production within polluted agroecosystems.

Formation of partially embedded Au nanostructures: Ion beam irradiation on thin film.

The Au partially embedded nanostructure (PEN) is synthesized by ion irradiation on an Au thin film deposited on a glass substrate using a 50 keV Ar ion. Scanning electron microscopy results show ion beam-induced restructuring from irregularly shaped nanostructures (NSs) to spherical Au NSs, and further ion irradiation leads to the formation of well-separated spherical nanoparticles. Higuchi's algorithm of surface analysis is utilized to find the evolution of surface morphology with ion irradiation in terms of the Hurst exponent and fractal dimension. The Au PEN is evidenced by Rutherford backscattering spectrometry and optical studies. Also, the depth of the mechanism behind synthesized PEN is explained on the basis of theoretical simulations, namely, a unified thermal spike and a Monte Carlo simulation consisting of dynamic compositional changes (TRIDYN). Another set of plasmonic NSs was formed on the surface by thermal annealing of the Au film on the substrate. Glucose sensing has been studied on the two types of plasmonic layers: nanoparticles on the surface and PEN. The results reveal the sensing responses of both types of plasmonic layers. However, PEN retains its plasmonic behavior as the NSs are still present after washing with water, which demonstrates the potential for reusability. RESEARCH HIGHLIGHTS: Synthesis of PENs by ion irradiation Utilization of Higuchi's algorithm to explore the surface morphology. Unified thermal spike and TRIDYN simulations being used to explain the results. Glucose is only used as a test case for reusability of substrate.

Metabolomics-Based Mechanistic Insights into Revealing the Adverse Effects of Pesticides on Plants: An Interactive Review.

In plant biology, metabolomics is often used to quantitatively assess small molecules, metabolites, and their intermediates in plants. Metabolomics has frequently been applied to detect metabolic alterations in plants exposed to various biotic and abiotic stresses, including pesticides. The widespread use of pesticides and agrochemicals in intensive crop production systems is a serious threat to the functionality and sustainability of agroecosystems. Pesticide accumulation in soil may disrupt soil-plant relationships, thereby posing a pollution risk to agricultural output. Application of metabolomic techniques in the assessment of the biological consequences of pesticides at the molecular level has emerged as a crucial technique in exposome investigations. State-of-the-art metabolomic approaches such as GC-MS, LC-MS/MS UHPLC, UPLC-IMS-QToF, GC/EI/MS, MALDI-TOF MS, and 1H-HR-MAS NMR, etc., investigating the harmful effects of agricultural pesticides have been reviewed. This updated review seeks to outline the key uses of metabolomics related to the evaluation of the toxicological impacts of pesticides on agronomically important crops in exposome assays as well as bench-scale studies. Overall, this review describes the potential uses of metabolomics as a method for evaluating the safety of agricultural chemicals for regulatory applications. Additionally, the most recent developments in metabolomic tools applied to pesticide toxicology and also the difficulties in utilizing this approach are discussed.

Zinc-solubilizing Bacillus spp. in conjunction with chemical fertilizers enhance growth, yield, nutrient content, and zinc biofortification in wheat crop.

Micronutrient deficiency is a serious health issue in resource-poor human populations worldwide, which is responsible for the death of millions of women and underage children in most developing countries. Zinc (Zn) malnutrition in middle- and lower-class families is rampant when daily calorie intake of staple cereals contains extremely low concentrations of micronutrients, especially Zn and Fe. Looking at the importance of the problem, the present investigation aimed to enhance the growth, yield, nutrient status, and biofortification of wheat crop by inoculation of native zinc-solubilizing Bacillus spp. in conjunction with soil-applied fertilizers (NPK) and zinc phosphate in saline soil. In this study, 175 bacterial isolates were recovered from the rhizosphere of wheat grown in the eastern parts of the Indo-Gangetic Plain of India. These isolates were further screened for Zn solubilization potential using sparingly insoluble zinc carbonate (ZnCO3), zinc oxide (ZnO), and zinc phosphate {Zn3(PO4)2} as a source of Zn under in vitro conditions. Of 175 bacterial isolates, 42 were found to solubilize either one or two or all the three insoluble Zn compounds, and subsequently, these isolates were identified based on 16S rRNA gene sequences. Based on zone halo diameter, solubilization efficiency, and amount of solubilized zinc, six potential bacterial strains, i.e., Bacillus altitudinis AJW-3, B. subtilis ABW-30, B. megaterium CHW-22, B. licheniformis MJW-38, Brevibacillus borstelensis CHW-2, and B. xiamenensis BLW-7, were further shortlisted for pot- and field-level evaluation in wheat crop. The results of the present investigation clearly indicated that these inoculants not only increase plant growth but also enhance the yield and yield attributes. Furthermore, bacterial inoculation also enhanced available nutrients and microbial activity in the wheat rhizosphere under pot experiments. It was observed that the application of B. megaterium CHW-22 significantly increased the Zn content in wheat straw and grains along with other nutrients (N, P, K, Fe, Cu, and Mn) followed by B. licheniformis MJW-38 as compared to other inoculants. By and large, similar observations were recorded under field conditions. Interestingly, when comparing the nutrient use efficiency (NUE) of wheat, bacterial inoculants showed their potential in enhancing the NUE in a greater way, which was further confirmed by correlation and principal component analyses. This study apparently provides evidence of Zn biofortification in wheat upon bacterial inoculation in conjunction with chemical fertilizers and zinc phosphate in degraded soil under both nethouse and field conditions.

Bacterial ACC deaminase: Insights into enzymology, biochemistry, genetics, and potential role in amelioration of environmental stress in crop plants.

Growth and productivity of crop plants worldwide are often adversely affected by anthropogenic and natural stresses. Both biotic and abiotic stresses may impact future food security and sustainability; global climate change will only exacerbate the threat. Nearly all stresses induce ethylene production in plants, which is detrimental to their growth and survival when present at higher concentrations. Consequently, management of ethylene production in plants is becoming an attractive option for countering the stress hormone and its effect on crop yield and productivity. In plants, ACC (1-aminocyclopropane-1-carboxylate) serves as a precursor for ethylene production. Soil microorganisms and root-associated plant growth promoting rhizobacteria (PGPR) that possess ACC deaminase activity regulate growth and development of plants under harsh environmental conditions by limiting ethylene levels in plants; this enzyme is, therefore, often designated as a "stress modulator." TheACC deaminase enzyme, encoded by the AcdS gene, is tightly controlled and regulated depending upon environmental conditions. Gene regulatory components of AcdS are made up of the LRP protein-coding regulatory gene and other regulatory components that are activated via distinct mechanisms under aerobic and anaerobic conditions. ACC deaminase-positive PGPR strains can intensively promote growth and development of crops being cultivated under abiotic stresses including salt stress, water deficit, waterlogging, temperature extremes, and presence of heavy metals, pesticides and other organic contaminants. Strategies for combating environmental stresses in plants, and improving growth by introducing the acdS gene into crop plants via bacteria, have been investigated. In the recent past, some rapid methods and cutting-edge technologies based on molecular biotechnology and omics approaches involving proteomics, transcriptomics, metagenomics, and next generation sequencing (NGS) have been proposed to reveal the variety and potential of ACC deaminase-producing PGPR that thrive under external stresses. Multiple stress-tolerant ACC deaminase-producing PGPR strains have demonstrated great promise in providing plant resistance/tolerance to various stressors and, therefore, it could be advantageous over other soil/plant microbiome that can flourish under stressed environments.

The Exogenous Application of 24-Epibrassinolide (24-EBL) Increases the Cd and Pb Resilience in Zea mays (L.) by Regulating the Growth and Physiological Mechanism.

Heavy metals (HMs) at a concentration above the threshold level act as environmental pollutants and very often threaten the agricultural productivity globally. Finding affordable and environmentally sustainable deliverables to address this issue is therefore a top focus. Phytohormones alleviate the HMs-induced toxicity and positively influence the plant growth. Considering the importance of phytohormones, the present study aimed to assess the effect of 24-epibrassinolide (24-EBL; 10 µM) as seed soaking treatment on growth performance of Zea mays (L.) contaminated separately with increasing concentrations (50-400 mg.kg-1) of lead (Pb) and cadmium (Cd). With increasing metal concentrations, growth and plant biometric criteria were reduced. For instance, Cd at 400 mg.kg-1 soil reduced the germination efficiency (56%), root (77%) and shoot (69%) dry weight, total chlorophyll (64%), and carotenoid content (45%). Contrarily, both HMs caused increase in stress biomarkers and antioxidant enzymes in seedling. However, exogenous administration of 24-EBL significantly enhanced the growth attributes, photosynthetic pigments, proline, MDA, and antioxidant enzyme activity while reducing the harmful effects of HMs stress on Z. mays. For instance, 24-EBL (10 µM) improved the germination percentage, root biomass, chl a, chl b, total chlorophyll, and carotenoid content by 16, 21, 17, 34, 18, and 15%, respectively, in 50 mg.Pb.kg-1 soil-treated Z. mays plants. Furthermore, the amounts of proline, MDA, and antioxidant enzymes in foliage of Z. mays were interestingly and dramatically lowered by 24-EBL application. Uptake of metals in plant organs was significantly reduced when 24-EBL was applied to Pb- and Cd-treated Z. mays. The recent findings help us better understand how 24-EBL regulates growth and development of Z. mays as well as how it boosts HMs' resilience, which could increase the possibility of employing 24-EBL to increase Z. mays productivity. Thus, the present findings confirmed the potentiality of pre-soaking the seed in 24-EBL solution that neutralizes the toxic effects of heavy metals in Z. mays plants. Therefore, it is suggested that applying phytohormones including 24-EBL in removal of heavy metal stress in plants is the best possible solution in sustainable agriculture.

Arbuscular mycorrhizal fungi-mediated activation of plant defense responses in direct seeded rice (Oryza sativa L.) against root-knot nematode Meloidogyne graminicola.

Rhizosphere is the battlefield of beneficial and harmful (so called phytopathogens) microorganisms. Moreover, these microbial communities are struggling for their existence in the soil and playing key roles in plant growth, mineralization, nutrient cycling and ecosystem functioning. In the last few decades, some consistent pattern have been detected so far that link soil community composition and functions with plant growth and development; however, it has not been studied in detail. AM fungi are model organisms, besides potential role in nutrient cycling; they modulate biochemical pathways directly or indirectly which lead to better plant growth under biotic and abiotic stress conditions. In the present investigations, we have elucidated the AM fungi-mediated activation of plant defense responses against Meloidogyne graminicola causing root-knot disease in direct seeded rice (Oryza sativa L.). The study describes the multifarious effects of Funneliformis mosseae, Rhizophagus fasciculatus, and Rhizophagus intraradices inoculated individually or in combination under glasshouse conditions in rice plants. It was found that F. mosseae, R. fasciculatus and R. intraradices when applied individually or in combination modulated the biochemical and molecular mechanisms in the susceptible and resistant inbred lines of rice. AM inoculation significantly increased various plant growth attributes in plants with simultaneous decrease in the root-knot intensity. Among these, the combined application of F. mosseae, R. fasciculatus, and R. intraradices was found to enhance the accumulation and activities of biomolecules and enzymes related to defense priming as well as antioxidation in the susceptible and resistant inbred lines of rice pre-challenged with M. graminicola. The application of F. mosseae, R. fasciculatus and R. intraradices, induced the key genes involved in plant defense and signaling and it has been demonstrated for the first time. Results of the present investigation advocated that the application of F. mosseae, R. fasciculatus and R. intraradices, particularly a combination of all three, not only helped in the control of root-knot nematodes but also increased plant growth as well as enhances the gene expression in rice. Thus, it proved to be an excellent biocontrol as well as plant growth-promoting agent in rice even when the crop is under biotic stress of the root-knot nematode, M. graminicola.

Unraveling the mechanism of sulfur nutrition in pigeonpea inoculated with sulfur-oxidizing bacteria.

An investigation was carried out to understand the mechanism(s) involved in the uptake of sulfur (S) as sulfate in pigeonpea following single inoculation of two sulfur-oxidizing bacteria (SOB), Stenotrophomonas maltophilia and Stenotrophomonas pavanii in the treatments amended with either elemental sulfur (S0) or sulfate (S6). Colonization potential and biofilm formation were analyzed through confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM). Furthermore, the effect of seed inoculation on root architecture, expression of genes involved in sulfur oxidation (sox) in bacterial inoculants, and genes involved in sulfate transport in pigeonpea (PpSULTR) were analyzed to correlate with the higher uptake of S in roots and shoots of pigeonpea. Both the SOB exhibited a good colonization potential and biofilm formation on the roots of pigeonpea. Among the 11 sox genes targeted in rhizosphere of pigeonpea, expression was achieved for seven genes, which showed 2-fold increase in treatments inoculated with S. maltophilia and amended with either S6 or S0. The inoculation of S. maltophilia and amendment of S0 led to increased expression of PpSULTR genes by several folds in roots. The inoculation of SOB had a significant influence on non-enzymatic (osmolytes like proline) and enzymatic (PAL, peroxidase, superoxide dismutase, and catalase) levels. The results revealed a significant increase in sulfur uptake in roots and shoots in treatment inoculated with S. maltophilia and amended with S6. The investigation showed that the SOB-mediated over-expression of PpSULTR genes in roots of pigeonpea and sox genes in the rhizosphere were acting synergistically in facilitating higher uptake and translocation of S in roots and shoots of pigeonpea plants.

Stress-Tolerant Endophytic Isolate Priestia aryabhattai BPR-9 Modulates Physio-Biochemical Mechanisms in Wheat (Triticum aestivum L.) for Enhanced Salt Tolerance.

In efforts to improve plant productivity and enhance defense mechanisms against biotic and abiotic stresses, endophytic bacteria have been used as an alternative to chemical fertilizers and pesticides. In the current study, 25 endophytic microbes recovered from plant organs of Triticum aestivum L. (wheat) were assessed for biotic (phyto-fungal pathogens) and abiotic (salinity, drought, and heavy metal) stress tolerance. Among the recovered isolates, BPR-9 tolerated maximum salinity (18% NaCl), drought (15% PEG-6000), and heavy metals (µg mL-1): Cd (1200), Cr (1000), Cu (1000), Pb (800), and Hg (30). Based on phenotypic and biochemical characteristics, as well as 16S rDNA gene sequencing, endophytic isolate BPR-9 was recognized as Priestia aryabhattai (accession no. OM743254.1). This isolate was revealed as a powerful multi-stress-tolerant crop growth promoter after extensive in-vitro testing for plant growth-promoting attributes, nutrient (phosphate, P; potassium, K; and zinc, Zn) solubilization efficiency, extracellular enzyme (protease, cellulase, amylase, lipase, and pectinase) synthesis, and potential for antagonistic activity against important fungal pathogens viz. Alternaria solani, Rhizoctonia solani, Fusarium oxysporum, and Ustilaginoidea virens. At elevated salt levels, increases were noted in indole-3-acetic acid; siderophores; P, K, and Zn-solubilization; ACC deaminase; and ammonia synthesized by Priestia aryabhattai. Additionally, under in-vitro plant bioassays, wheat seedlings inoculated with P. aryabhattai experienced superior growth compared to non-inoculated seedlings in high salinity (0-15% NaCl) environment. Under NaCl stress, germination rate, plant length, vigor indices, and leaf pigments of wheat seedlings significantly increased following P. aryabhattai inoculation. Furthermore, at 2%-NaCl, B. aryabhattai greatly and significantly (p ≤ 0.05) decreased relative leaf water content, membrane damage, and electrolyte leakage compared with the non-inoculated control. Catalase, superoxide dismutase, and peroxidase activity increased by 29, 32, and 21%, respectively, in wheat seedlings exposed to 2% NaCl and inoculated with the bacteria. The present findings demonstrate that endophytic P. aryabhattai strains might be used in the future as a multi-stress reducer and crop growth promoter in agronomically important crops including cereals.

Endophytic Bacillus subtilis antagonize soil-borne fungal pathogens and suppress wilt complex disease in chickpea plants (Cicer arietinum L.).

The present study aimed to identify potential endophytic bacteria antagonistic against three soil-borne fungal pathogens, Rhizoctonia solani, Sclerotium rolfsii, and Fusarium oxysporum f.sp. ciceri causing root rot, collar rot, and fungal wilt diseases in chickpea plants, respectively. A total of 255 bacterial endophytes were isolated from the leaves, stems, and roots of seven different crop plants (chickpea, tomato, wheat, berseem, mustard, potato, and green pea). The dual culture-based screening for antifungal properties indicated that three endophytic isolates had strong inhibition (>50%) against all three pathogens tested. Based on morphological, biochemical, and molecular characterization, the selected isolates (TRO4, CLO5, and PLO3) were identified as different strains of Bacillus subtilis. The bacterial endophytes (TRO4 and CLO5) were positive for plant growth promoting (PGP) traits viz., ammonia, siderophore, and indole-3-acetic acid (IAA) production. The bio-efficacy of the endophytes (TRO4, CLO5, and PLO3) was tested by an in planta trial in chickpea pre-challenged with R. solani, S. rolfsii, and F. oxysporum f.sp. ciceri. The B. subtilis strains TRO4 and CLO5 were found to be effective in reducing percent disease incidence (p ≤ 0.05) and enhancing plant growth parameters. The different root parameters viz. root length (mm), surface area (cm2), root diameter (mm), and root volume (cm3) were significantly (p ≤ 0.05) increased in TRO4 and CLO5 inoculated chickpea plants. Confocal Scanning Laser Microscopy showed heavy colonization of bacteria in the roots of endophyte-inoculated chickpea plants. The inoculation of endophytic Bacillus subtilis strains TRO4 and CLO5 in chickpea plants through seed biopriming reduced the accumulation of superoxide, enhanced the plant defense enzymes, and induced the expression of Pathogenesis-Related (PR) genes. Semi-quantitative analysis of defense-related genes showed differential activation of PR genes (60srp and IFR) by endophyte inoculation. The results of the present study reveal the antagonistic potential of B. subtilis strains TRO4 and CLO5 against three major soil-borne fungal pathogens and their ability to suppress wilt complex disease in chickpea plants. This is the first report on the simultaneous suppression of three major soil-borne fungal pathogens causing wilt complex in chickpea plants by endophytic B. subtilis strains.

A Comparative Analysis of Microbe-Based Technologies Developed at ICAR-NBAIM Against Erysiphe necator Causing Powdery Mildew Disease in Grapes (Vitis vinifera L.).

Globally, Erysiphe necator causing powdery mildew disease in grapevines (Vitis vinifera L.) is the second most important endemic disease, causing huge economic losses every year. At present, the management of powdery mildew in grapes is largely dependent upon the use of chemical fungicides. Grapes are being considered as one of the high pesticide-demanding crops. Looking at the residual impact of toxic chemical pesticides on the environment, animal, and human health, microbe-based strategies for control of powdery mildew is an emerging technique. It offers an environment-friendly, residue-free, and effective yet safer approach to control powdery mildew disease in grapes. The mode of action is relatively diverse as well as specific to different pathosystems. Hence, the aim of this study was to evaluate the microbe-based technologies, i.e., Eco-pesticide®, Bio-Pulse®, and Bio-Care 24® developed at the Plant-Microbe Interaction and Rhizosphere Biology Lab, ICAR-NBAIM, Kushmaur, against grape powdery mildew and to integrate these technologies with a safer fungicide (sulfur) to achieve better disease control under organic systems of viticulture. The experiments were conducted at four different locations, namely, the vineyards of ICAR-NRCG, Rajya Draksha Bagayatdar Sangh (MRDBS), and two farmers' fields at Narayangaon and Junnar in the Pune district of Maharashtra. A significantly lower percent disease index (PDI) was recorded on the leaves of grape plants treated with Eco-Pesticide®/sulfur (22.37) followed by Bio-Pulse®/sulfur (22.62) and Bio-Care 24®/sulfur (24.62) at NRCG. A similar trend was observed with the lowest PDI on bunches of Eco-pesticide® /sulfur-treated plants (24.71) followed by Bio-Pulse®/sulfur (24.94) and Bio-Care®/sulfur (26.77). The application of microbial inoculants singly or in combination with sulfur has a significant positive impact on the qualitative parameters such as pH, total soluble solids (TSS), acidity, berry diameter, and berry length of the grapes at different locations. Among all the treatments, the Bio-Pulse®/sulfur treatment showed the highest yield per vine (15.02 kg), which was on par with the treatment Eco-Pesticide®/sulfur (14.94). When compared with the yield obtained from the untreated control, 2.5 to 3 times more yield was recorded in the plants treated with either of the biopesticides used in combination with sulfur. Even in the case of individual inoculation, the yield per vine was approximately two times higher than the untreated control and water-treated plants across the test locations. Results suggested that microbial technologies not only protect grapevines from powdery mildew but also enhance the quality parameters with increased yield across the test locations.

Modulation in Biofertilization and Biofortification of Wheat Crop by Inoculation of Zinc-Solubilizing Rhizobacteria.

Zinc is an important micronutrient needed for the optimum growth and development of plants. Contrary to chemical zinc fertilizers, the use of zinc-solubilizing bacteria is an environmentally friendly option for zinc enrichment in edible parts of crops. This study was conducted with the objective of selecting potential zinc-solubilizing rhizobacteria from the rhizosphere of chickpea grown in soils of eastern Uttar Pradesh and further assessing their impact on the magnitude of zinc assimilation in wheat crops. Among 15 isolates, CRS-9, CRS-17, CRS-30, and CRS-38 produced net soluble zinc in broth to the tune of 6.1, 5.9, 5.63, and 5.6 µg ml-1, respectively, in zinc phosphate with the corresponding pH of 4.48, 5.31, 5.2, and 4.76. However, the bacterial strains CRS-17, CRS-30, CRS-38, and CRS-9 showed maximum zinc phosphate solubilization efficiency of 427.79, 317.39, 253.57, and 237.04%, respectively. The four bacterial isolates were identified as Bacillus glycinifermentans CRS-9, Microbacterium oxydans CRS-17, Paenarthrobacter nicotinovorans CRS-30, and Bacillus tequilensis CRS-38 on the basis of morphological and biochemical studies and 16S rRNA gene sequencing. Bacterial inoculants significantly colonized the roots of wheat plants and formed a biofilm in the root matrix. These strains significantly increased seed germination (%) and vigor indices in wheat grown under glasshouse conditions. After 30 days of sowing of wheat under microcosm conditions, eight zinc transporter (TaZIP) genes were expressed maximally in roots, with concomitant accumulation of higher zinc content in the bacterially treated plant compared to the absolute control. Out of the four strains tested, two bacteria, B. tequilensis CRS-38 and P. nicotinovorans CRS-30, improved seed germination (%), vigor indices (2-2.5 folds), plant biomass, grain yield (2.39 g plant-1), and biofortificated grains (54.25 µg g-1Zn) of wheat. To the best of our knowledge, this may be the first report on the presence of zinc solubilization trait in B. glycinifermentans CRS-9, M. oxydans CRS-17, and P. nicotinovorans CRS-30.

Bio-priming with a consortium of Streptomyces araujoniae strains modulates defense response in chickpea against Fusarium wilt.

Wilt caused by Fusarium oxysporum f. sp. ciceris (Foc) is one of the major diseases of chickpea affecting the potential yield significantly. Productivity and biotic stress resilience are both improved by the association and interaction of Streptomyces spp. with crop plants. In the present study, we evaluated two Streptomyces araujoniae strains (TN11 and TN19) for controlling the wilt of chickpea individually and as a consortium. The response of Foc challenged chickpea to inoculation with S. araujoniae TN11 and TN19 individually and as a consortium was recorded in terms of changes in physio-biochemical and expression of genes coding superoxide dismutase (SOD), peroxidase, and catalase. Priming with a consortium of TN11 and TN19 reduced the disease severity by 50-58% when challenged with Foc. Consortium primed-challenged plants recorded lower shoot dry weight to fresh weight ratio and root dry weight to fresh weight ratio as compared to challenged non-primed plants. The pathogen-challenged consortium primed plants recorded the highest accumulation of proline and electrolyte leakage. Similarly, total chlorophyll and carotenoids were recorded highest in the consortium treatment. Expression of genes coding SOD, peroxidase, and catalase was up-regulated which corroborated with higher activities of SOD, peroxidase, and catalase in consortium primed-challenged plants as compared to the challenged non-primed plants. Ethyl acetate extracts of TN11 and TN19 inhibited the growth of fungal pathogens viz., Fusarium oxysporum f. sp. ciceris. Macrophomina phaseolina, F. udum, and Sclerotinia sclerotiarum by 54-73%. LC-MS analyses of the extracts showed the presence of a variety of antifungal compounds like erucamide and valinomycin in TN11 and valinomycin and dinactin in TN19. These findings suggest that the consortium of two strains of S. araujoniae (TN11 and TN19) can modulate defense response in chickpea against wilt and can be explored as a biocontrol strategy.

Nano-pesticidal potential of Cassia fistula (L.) leaf synthesized silver nanoparticles (Ag@CfL-NPs): Deciphering the phytopathogenic inhibition and growth augmentation in Solanum lycopersicum (L.).

Plant-based synthesis of silver nanoparticles (Ag-NPs) has emerged as a potential alternative to traditional chemical synthesis methods. In this context, the aim of the present study was to synthesize Ag-NPs from Cassia fistula (L.) leaf extract and to evaluate their nano-pesticidal potential against major phyto-pathogens of tomato. From the data, it was found that particle size of spherical C. fistula leaf synthesized (Ag@CfL-NPs) varied from 10 to 20 nm, with the average diameter of 16 nm. Ag@CfL-NPs were validated and characterized by UV-visible spectroscopy (surface resonance peak λ max = 430 nm), energy dispersive spectrophotometer (EDX), Fourier transform infrared (FTIR), and X-ray diffraction pattern (XRD), and electron microscopy; scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The FTIR spectra verified the participation of various living molecules (aromatic/aliphatic moieties and proteins) in synthesized Ag@CfL-NPs. The anti-phytopathogenic potential of Ag@CfL-NPs was assessed under in vitro conditions. Increasing doses of Ag@CfL-NPs exhibited an inhibitory effect against bacterial pathogen Pseudomonas syringae and 400 µg Ag@CfL-NPs ml-1 caused a reduction in cellular viability, altered bacterial morphology, and caused cellular death Furthermore, Ag@CfL-NPs reduced exopolysaccharides (EPS) production and biofilm formation by P. syringae Additionally, Ag@CfL-NPs showed pronounced antifungal activity against major fungal pathogens. At 400 µg Ag@CfL-NPs ml-1, sensitivity of tested fungi followed the order: Fusarium oxysporum (76%) > R. solani (65%) > Sarocladium (39%). Furthermore, 400 µg Ag@CfL-NPs ml-1 inhibited the egg-hatching and increased larval mortality of Meloidogyne incognita by 82 and 65%, respectively, over control. Moreover, pot studies were performed to assess the efficacy of Ag@CfL-NPs to phyto-pathogens using tomato (Solanum lycopersicum L.) as a model crop. The applied phyto-pathogens suppressed the biological, physiological, and oxidative-stress responsiveness of tomatoes. However, 100 mg Ag@CfL-NPs kg-1 improved overall performance and dramatically increased the root length, dry biomass, total chlorophyll, carotenoid, peroxidase (POD), and phenylalanine ammonia lyase (PAL) activity over pathogens-challenged tomatoes. This study is anticipated to serve as an essential indication for synthesis of efficient nano-control agents, which would aid in the management of fatal phyto-pathogens causing significant losses to agricultural productivity. Overall, our findings imply that Ag@CfL-NPs as nano-pesticides might be used in green agriculture to manage the diseases and promote plant health in a sustainable way.