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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.

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.

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 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.

The present investigation was carried out to isolate, identify, and characterize sulfur-oxidizing bacteria (SOB) from coal mines and to evaluate the efficient strains for their ability to influence plant growth and S uptake in pigeonpea. Thirteen bacterial isolates belonging to Stenotrophomonas maltophilia (2), Stenotrophomonas pavanii (2), Rhizobium pusense (5), Bacillus velezensis (2), and Paenibacillus massiliensis (2) were obtained. Among these, seven strains that could reduce the pH of thiosulfate broth were further characterized for sulfur oxidation, plant growth-promoting (PGP) attributes, and in planta studies. Among the seven strains characterized, maximum sulfate ion was recorded for S. maltophilia DRC-18-7A (311.43 mg L−1) closely followed by S. pavanii DRC-18-7B (273.44 mg L−1) and S. maltophilia DRC-18-10 (265.75 mg L−1) after 21 days of inoculation. Among the PGP attributes quantified, maximum P solubilization was recorded in case of S. maltophilia DRC-18-7A (24.39 μg ml−1), while highest siderophore production and IAA production were recorded in S. maltophilia DRC-18-10 (14.25%) and R. pusense DRC-18-25 (15.21 μg ml−1), respectively. S. maltophilia DRC-18-7A closely followed by S. pavanii DRC-18-7B outperformed others in enhancing seed germination (%) and vigour indices. Results clearly indicated that microbial inoculants colonized the plant roots and developed biofilm on the root surface. It was further observed that plants treated with microbial inoculants induce an early formation of secondary and tertiary roots in the pigeonpea compared to the untreated control which was further confirmed by assessing the root architecture using the root scanner. Inoculation of these two strains to pigeonpea significantly enhanced plant growth parameters, the activity of reactive oxygen scavenging (ROS) enzymes, and accumulation of flavonoids, carotenoids, and proline both under sterilized and non-sterilized growth medium (sand and soil in 1:3 ratio). The application of microbial inoculants significantly increased the uptake of nitrogen, phosphorus, potassium, and sulfur in plant shoots. Further, transcript level of phosphate, potassium, and sulfur transporter genes significantly increases upon microbial inoculation leading to increased uptake and translocation of P, K, and S in the pigeonpea. The results indicate that S. maltophilia DRC-18-7A and S. pavanii DRC-18-7B could be recommended as inoculants for pigeonpea to improve its growth and sulfur nutrition.

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.

Endophytic bacilli of ethano-botanical plant Ocimum tenuiflorum were screened for salt stress-alleviating traits in tomato. Four promising O. tenuiflorum endophytes (Bacillus safensis BTL5, Bacillus haynesii GTR8, Bacillus paralicheniformis GTR11, and Bacillus altitudinis GTS16) were used in this study. Confocal scanning laser microscopic studies revealed the inter-genera colonization of O. tenuiflorum endophytes in tomato plants, giving insights for widening the applicability of potential endophytes to other crops. Furthermore, in a pot trial under 150 mM NaCl concentration, the inoculated endophytes contributed in reducing salt toxicity and improving recovery from salt-induced oxidative stress by different mechanisms. Reduction in reactive oxygen species (ROS) (sub-cellular H2O2 and superoxide) accumulation was observed besides lowering programmed cell death and increasing chlorophyll content. Endophyte inoculation supplemented the plant antioxidant enzyme system via the modulation of enzymatic antioxidants, viz., peroxidase, ascorbate peroxidase, superoxide dismutase, and catalase, apart from increasing proline and total phenolics. Antioxidants like proline have dual roles of antioxidants and osmoregulation, which might also have contributed to improved water relation under elevated salinity. Root architecture, viz., root length, projection area, surface area, average diameter, tips, forks, crossings, and the number of links, was improved upon inoculation, indicating healthy root growth and enhanced nutrient flow and water homeostasis. Regulation of Na+/K+ balance and water homeostasis in the plants were also evident from the modulation in the expression of abiotic stress-responsive genes, viz., LKT1, NHX1, SOS1, LePIP2, SlERF16, and SlWRKY39. Shoot tissues staining with light-excitable Na+ indicator Sodium GreenTM Tetra (tetramethylammonium) salt showed low sodium transport and accumulation in endophyte-inoculated plants. All four endophytes exhibited different mechanisms for stress alleviation and indicated complementary effects on plant growth. Furthermore, this could be harnessed in the form of a consortium for salt stress alleviation. The present study established inter-genera colonization of O. tenuiflorum endophytes in tomato and revealed its potential in maintaining Na+/K+ balance, reducing ROS, and improving root architecture under elevated salinity.

Spot blotch disease of wheat, caused by the fungus Bipolaris sorokiniana (Sacc.) Shoem., produces several toxins which interact with the plants and thereby increase the blightening of the wheat leaves, and Bipolaroxin is one of them. There is an urgent need to decipher the molecular interaction between wheat and the toxin Bipolaroxin for in-depth understanding of host–pathogen interactions. In the present study, we have developed the three-dimensional structure of G-protein alpha subunit from Triticum aestivum. Molecular docking studies were performed using the active site of the modeled G-protein alpha and cryo-EM structure of beta subunit from T. aestivum and ‘Bipolaroxin’. The study of protein–ligand interactions revealed that six H-bonds are mainly formed by Glu29, Ser30, Lys32, and Ala177 of G-alpha with Bipolaroxin. In the beta subunit, the residues of the core beta strand domain participate in the ligand interaction where Lys256, Phe306, and Leu352 formed seven H-bonds with the ligand Bipolaroxin. All-atoms molecular dynamics (MD) simulation studies were conducted for G-alpha and -beta subunit and Bipolaroxin complexes to explore the stability, conformational flexibility, and dynamic behavior of the complex system. In planta studies clearly indicated that application of Bipolaroxin significantly impacted the physio-biochemical pathways in wheat and led to the blightening of leaves in susceptible cultivars as compared to resistant ones. Further, it interacted with the Gα and Gβ subunits of G-protein, phenylpropanoid, and MAPK pathways, which is clearly supported by the qPCR results. This study gives deeper insights into understanding the molecular dialogues between Bipolaroxin and the Gα and Gβ subunits of the wheat heterotrimeric G-protein during host–pathogen interaction.

Salt stress hampers plant growth and development. It is now becoming one of the most important threats to agricultural productivity. Rhizosphere microorganisms play key roles in modulating cellular responses and enable plant tolerant to salt stress, but the detailed mechanisms of how this occurs need in-depth investigation. The present study elucidated that the microbe-mediated restructuring of the cellular responses leads to ecological fitness and adaptiveness to the maize (Zea mays L.) grown in saline–sodic soil. In the present study, effects of seed biopriming with B. safensis MF-01, B. altitudinis MF-15, and B. velezensis MF-08 singly and in consortium on different growth parameters were recorded. Soil biochemical and enzymatic analyses were performed. The activity and gene expression of High-Affinity K+ Transporter (ZmHKT-1), Sodium/Hydrogen exchanger 1 (zmNHX1), and antioxidant enzymes (ZmAPX1.2, ZmBADH-1, ZmCAT, ZmMPK5, ZmMPK7, and ZmCPK11) were studied. The expression of genes related to lateral root development (ZmHO-1, ZmGSL-1, and ZmGSL-3) and root architecture were also carried out. Seeds bioprimed with consortium of all three strains have been shown to confer increased seed germination (23.34–26.31%) and vigor indices (vigor index I: 38.71–53.68% and vigor index II: 74.11–82.43%) as compared to untreated control plant grown in saline–sodic soil at 30 days of sowing. Results indicated that plants treated with consortium of three strains induced early production of adventitious roots (tips: 4889.29, forks: 7951.57, and crossings: 2296.45) in maize compared to plants primed with single strains and untreated control (tips: 2019.25, forks: 3021.45, and crossings: 388.36), which was further confirmed by assessing the transcript level of ZmHO-1 (7.20 folds), ZmGSL-1 (4.50 folds), and ZmGSL-3 (12.00 folds) genes using the qPCR approach. The uptake and translocation of Na+, K+, and Ca2+ significantly varied in the plants treated with bioagents alone or in consortium. qRT-PCR analysis also revealed that the ZmHKT-1 and zmNHX1 expression levels varied significantly in the maize root upon inoculation and showed a 6- to 11-fold increase in the plants bioprimed with all the three strains in combination. Further, the activity and gene expression levels of antioxidant enzymes were significantly higher in the leaves of maize subjected seed biopriming with bioagents individually or in combination (3.50- to 12.00-fold). Our research indicated that ZmHKT-1 and zmNHX1 expression could effectively enhance salt tolerance by maintaining an optimal Na+/K+ balance and increasing the antioxidant activity that keeps reactive oxygen species at a low accumulation level. Interestingly, up-regulation of ZmHKT-1, NHX1, ZmHO-1, ZmGSL-1, and ZmGSL-3 and genes encoding antioxidants regulates the cellular responses that could effectively enhance the adaptiveness and ultimately leads to better plant growth and grain production in the maize crop grown in saline–sodic soil.

Purpose: The study was aimed at finding out the present pattern of referrals to the Pediatric Ophthalmology outpatient department (OPD) in a tertiary eye care hospital and thus identify the discrepancy of referral, if any, which would help to modify and enhance the practice guidelines. Methods: The study was conducted by retrospectively collecting data from all referral letters that were already uploaded in the Electronic Medical Report (EMR) against all patients from June 2019 to December 2019. All pediatric patients in the age group of 0–16 years were included in the study. The practicing field of referring clinicians was noted along with the maximum information that could be collected from the referral letter and were thus assessed for the quality, accuracy, and timely referral. Results: Out of 77 referrals received in the study period, six referral letters neither had any mention of the designation of the referring clinician nor any specific diagnosis or details. Thus, only 71 patients were included for further study. The referring clinicians were mainly ophthalmologists, pediatricians, general practitioners (GPs), and others (cardiologists, neurologists). Maximum patients were referred by ophthalmologists (76%) but visual acuity was noted only for 30% of these patients. Almost half of the referral diagnosis was accurate. Pediatrician referrals were found to be more detailed and précised. Conclusion: There is a need for a standardized hospital-specific format of referrals and basic training to primary care providers on some simple tests (Lights reflex tests) for identifying the “red flags” in pediatric eye examination and thus enhancing the quality and timely referral per se.