Bismuth sulfide micro flowers decorated nickel foam as a promising electrochemical sensor for quantitative analysis of melamine in bottled milk samples.

Here, we demonstrate hydrothermally grown bismuth sulfide (Bi2S3) micro flowers decorated nickel foam (NF) for electrochemical detection of melamine in bottled milk samples. The orthorhombic phase of hydrothermally grown Bi2S3is confirmed by the detailed characterization of x-ray diffraction and its high surface area micro flowers-like morphology is investigated via field emission scanning electron microscope. Furthermore, the surface chemical oxidation state and binding energy of Bi2S3/NF micro flowers is analyzed by x-ray photoelectron spectroscopy studies. The sensor exhibits a wide linear range of detection from 10 ng l-1to 1 mg l-1and a superior sensitivity of 3.4 mA cm-2to melamine using differential pulse voltammetry technique, with a lower limit of detection (7.1 ng l-1). The as-fabricated sensor is highly selective against interfering species of p-phenylenediamine (PPDA), cyanuric acid (CA), aniline, ascorbic acid, glucose (Glu), and calcium ion (Ca2+). Real-time analysis done in milk by the standard addition method shows an excellent recovery percentage of Ì´ 98%. The sensor's electrochemical mechanism studies reveal that the high surface area bismuth sulfide micro flowers surface interacts strongly with melamine molecules through hydrogen bonding and van der Waals forces, resulting in a significant change in the sensor's electrical properties while 3D skeletal Nickel foam as a substrate provides stability, enhances its catalytic activity by providing a more number /of active sites and facilitates rapid electron transfer. The work presented here confirms Bi2S3/NF as a high-performance electrode that can be used for the detection of other biomolecules used in clinical diagnosis and biomedical research.

Synergistic integration of Ni-metal organic framework/SnS2nanocomposite and nickel foam electrode for ultrasensitive and selective electrochemical detection of albumin in simulated human blood serum.

Albumin is a vital blood protein responsible for transporting metabolites and drugs throughout the body and serves as a potential biomarker for various medical conditions, including inflammatory, cardiovascular, and renal issues. This report details the fabrication of Ni-metal organic framework/SnS2nanocomposite modified nickel foam electrochemical sensor for highly sensitive and selective non enzymatic detection of albumin in simulated human blood serum samples. Ni-metal organic framework/SnS2nanocomposite was synthesized using solvothermal technique by combining Ni-metal-organic framework (MOF) with conductive SnS2leading to the formation of a highly porous material with reduced toxicity and excellent electrical conductivity. Detailed surface morphology and chemical bonding of the Ni-MOF/SnS2nanocomposite was studied using scanning electron microscopy, transmission electron microscopy, Fourier transform infra-red, and Raman analysis. The Ni-MOF/SnS2nanocomposite coated on Ni foam electrode demonstrated outstanding electrochemical performance, with a low limit of detection (0.44µM) and high sensitivity (1.3µA/pM/cm2) throughout a broad linear range (100 pM-10 mM). The remarkable sensor performance is achieved through the synthesis of a Ni-MOF/SnS2nanocomposite, enhancing electrocatalytic activity for efficient albumin redox reactions. The enhanced performance can be attributed due to the structural porosity of nickel foam and Ni-metal organic framework, which favours increased surface area for albumin interaction. The presence of SnS2shows stability in acidic and neutral solutions due to high surface to volume ratio which in turn improves sensitivity of the sensing material. The sensor exhibited commendable selectivity, maintaining its performance even when exposed to potential interfering substances like glucose, ascorbic acid, K+, Na+, uric acid, and urea. The sensor effectively demonstrates its accuracy in detecting albumin in real samples, showcasing substantial recovery percentages of 105.1%, 110.28%, and 91.16%.

Tin oxide-polyaniline nanocomposite modified nickel foam for highly selective and sensitive detection of cholesterol in simulated blood serum samples.

Cholesterol (CH) is a vital diagnostic marker for a variety of diseases, making its detection crucial in biological applications including clinical practice. In this work, we report the synthesis of tin oxide-polyaniline nanocomposite-modified nickel foam (SnO2-PANI/NF) for non-enzymatic detection of CH in simulated human blood serum. SnO2was synthesized via the hydrothermal method, followed by the synthesis of SnO2-PANI nanocomposite throughin situchemical polymerization of aniline using ammonium persulfate as the oxidizing agent. Morphological studies display agglomerated SnO2-PANI, which possess diameters ranging from an average particle size of ∼50 to ∼500 nm, and the XRD analysis revealed the tetragonal structure of the SnO2-PANI nanocomposite. Optimization studies demonstrating the effect of pH and weight percentage are performed to improve the electrocatalytic performance of the sensor. The non-enzymatic SnO2-PANI/NF sensor exhibits a linear range of 1-100µM with a sensitivity of 300µAµM-1/cm-2towards CH sensing and a low limit of detection of 0.25µM (=3 S m-1). SnO2-PANI/NF facilitates the electrooxidation of CH to form cholestenone by accepting electrons generated during the reaction and transferring them to the nickel foam electrode via Fe (III)/Fe (IV) conversion, resulting in an increased electrochemical current response. The SnO2-PANI/NF sensor demonstrated excellent selectivity against interfering species such as Na+, Cl-, K+, glucose, ascorbic acid, and SO42-. The sensor successfully determined the concentration of CH in simulated blood serum samples, demonstrating SnO2-PANI as a potential platform for a variety of electrochemical-based bioanalytical applications.

Metabacillus kandeliae sp. nov., isolated from the rhizosphere soil of a decayed mangrove plant Kandelia candel.

In this study, we isolated a novel Gram-stain-positive, aerobic, motile, rod-shaped bacterium, named GX 13764T, from the rhizosphere soil of a decayed mangrove plant Kandelia candel collected from Beihai, Guangxi, PR China, and characterized using a polyphasic taxonomic approach. The strain exhibited yellow-orange, round, convex, shiny, smooth, opaque and 2-3 mm diameter colonies on marine agar 2216 media after 3 days of incubation at 30 °C and was capable of growth at 4-45 °C (optimum, 30 °C), pH 5.0-9.0 (optimum, pH 7.0) and 0-4 % NaCl (w/v; optimum, 2 %). The strain was positive for catalase and negative for the oxidase. The main cellular fatty acid was anteiso-C15:0, iso-C15:0, iso-C16:0 and iso-C14:0. The cell-wall peptidoglycan comprised meso-diaminopimelic acid and the main menaquinone was MK-7. The polar lipids included one diphosphatidylglycerol, one phosphatidylglycerol, two glycolipids, two unidentified phospholipids and three unidentified lipids. Based on 16S rRNA gene analysis, GX 13764T presented the highest sequence similarity to Metabacillus mangrovi KCTC 33872T (97.04 %). The DNA G+C content of the type strain was 44.2 mol%. The average nucleotide identity values between GX 13764T and M. mangrovi KCTC 33872T, Metabacillus idriensis DSM 19097T and Metabacillus indicus LMG 22858T were 69.39, 68.87 and 68.95 %, respectively, with digital DNA-DNA hybridization values of 19.9, 19.5 and 19.5 %, respectively. Based on the polyphasic data, strain GX 13764T should be nominated as a novel species of the genus Metabacillus, for which the name Metabacillus kandeliae sp. nov. is proposed. The type strain is GX 13764T (=MCCC 1K06654T=KCTC 43366T).

Evaluating impacts of biogenic silver nanoparticles and ethylenediurea on wheat (Triticum aestivum L.) against ozone-induced damages.

Tropospheric ozone (O3) is a phytotoxic pollutant that leads to a reduction in crop yield. Nanotechnology offers promising solutions to stem such yield losses against abiotic stresses. Silver nanoparticles are major nanomaterials used in consumer products however, their impact on crops under abiotic stress is limited. In this study, we evaluated the anti-ozonant efficacy of biogenic silver nanoparticles (B-AgNPs) and compared them with a model anti-ozonant ethylenediurea (EDU) against ozone phyto-toxicity. Growth, physiology, antioxidant defense, and yield parameters in two wheat cultivars (HD-2967 & DBW-17), treated with B-AgNPs (25 mg/L and 50 mg/L) and EDU (150 mg/L and 300 mg/L), were studied at both vegetative and reproductive stages. During the experimental period, the average ambient ozone concentration and accumulated dose of ozone over a threshold of 40 ppb (AOT40) (8 h day-1) were found to be 60 ppb and 6 ppm h, respectively, which were sufficient to cause ozone-induced phyto-toxicity in wheat. Growth and yield for B-AgNPs as well as EDU-treated plants were significantly higher in both the tested cultivars over control ones. However, 25 mg/L B-AgNPs treatment showed a more pronounced effect in terms of yield attributes and its lower accumulation in grains for both cultivars. DBW-17 cultivar responded better with B-AgNPs and EDU treatments as compared to HD-2967. Meanwhile, foliar exposure of B-AgNPs (dose; 25 mg/L) significantly enhanced grain weight plant-1, thousand-grain weight, and harvest index by 54.22 %, 29.46 %, and 14.21 %, respectively in DBW-17, when compared to control. B-AgNPs could enhance ozone tolerance in wheat by increasing biochemical and physiological responses. It is concluded that B-AgNPs at optimum concentrations were as effective as EDU, hence could be a promising ozone protectant for wheat.

Unraveling Nitrogen Fixing Potential of Endophytic Diazotrophs of Different Saccharum Species for Sustainable Sugarcane Growth.

Sugarcane (Saccharum officinarum L.) is one of the world's highly significant commercial crops. The amounts of synthetic nitrogen (N2) fertilizer required to grow the sugarcane plant at its initial growth stages are higher, which increases the production costs and adverse environmental consequences globally. To combat this issue, sustainable environmental and economic concerns among researchers are necessary. The endophytic diazotrophs can offer significant amounts of nitrogen to crops through the biological nitrogen fixation mediated nif gene. The nifH gene is the most extensively utilized molecular marker in nature for studying N2 fixing microbiomes. The present research intended to determine the existence of novel endophytic diazotrophs through culturable and unculturable bacterial communities (EDBCs). The EDBCs of different tissues (root, stem, and leaf) of five sugarcane cultivars (Saccharum officinarum L. cv. Badila, S. barberi Jesw.cv Pansahi, S. robustum, S. spontaneum, and S. sinense Roxb.cv Uba) were isolated and molecularly characterized to evaluate N2 fixation ability. The diversity of EDBCs was observed based on nifH gene Illumina MiSeq sequencing and a culturable approach. In this study, 319766 operational taxonomic units (OTUs) were identified from 15 samples. The minimum number of OTUs was recorded in leaf tissues of S. robustum and maximum reads in root tissues of S. spontaneum. These data were assessed to ascertain the structure, diversity, abundance, and relationship between the microbial community. A total of 40 bacterial families with 58 genera were detected in different sugarcane species. Bacterial communities exhibited substantially different alpha and beta diversity. In total, 16 out of 20 genera showed potent N2-fixation in sugarcane and other crops. According to principal component analysis (PCA) and hierarchical clustering (Bray-Curtis dis) evaluation of OTUs, bacterial microbiomes associated with root tissues differed significantly from stem and leaf tissues of sugarcane. Significant differences often were observed in EDBCs among the sugarcane tissues. We tracked and validated the plethora of individual phylum strains and assessed their nitrogenase activity with a culture-dependent technique. The current work illustrated the significant and novel results of many uncharted endophytic microbial communities in different tissues of sugarcane species, which provides an experimental system to evaluate the biological nitrogen fixation (BNF) mechanism in sugarcane. The novel endophytic microbial communities with N2-fixation ability play a remarkable and promising role in sustainable agriculture production.

A Thermotolerant Marine Bacillus amyloliquefaciens S185 Producing Iturin A5 for Antifungal Activity against Fusarium oxysporum f. sp. cubense.

Fusarium wilt of banana (also known as Panama disease), is a severe fungal disease caused by soil-borne Fusarium oxysporum f. sp. cubense (Foc). In recent years, biocontrol strategies using antifungal microorganisms from various niches and their related bioactive compounds have been used to prevent and control Panama disease. Here, a thermotolerant marine strain S185 was identified as Bacillus amyloliquefaciens, displaying strong antifungal activity against Foc. The strain S185 possesses multiple plant growth-promoting (PGP) and biocontrol utility properties, such as producing indole acetic acid (IAA) and ammonia, assimilating various carbon sources, tolerating pH of 4 to 9, temperature of 20 to 50 °C, and salt stress of 1 to 5%. Inoculation of S185 colonized the banana plants effectively and was mainly located in leaf and root tissues. To further investigate the antifungal components, compounds were extracted, fractionated, and purified. One compound, inhibiting Foc with minimum inhibitory concentrations (MICs) of 25 µg/disk, was identified as iturin A5 by high-resolution electrospray ionization mass spectrometry (HR-ESI-MS) and nuclear magnetic resonance (NMR). The isolated iturin, A5, resulted in severe morphological changes during spore germination and hyphae growth of Foc. These results specify that B. amyloliquefaciens S185 plays a key role in preventing the Foc pathogen by producing the antifungal compound iturin A5, and possesses potential as a cost-effective and sustainable biocontrol strain for Panama disease in the future. This is the first report of isolation of the antifungal compound iturin A5 from thermotolerant marine B. amyloliquefaciens S185.

Diversity of nitrogen-fixing rhizobacteria associated with sugarcane: a comprehensive study of plant-microbe interactions for growth enhancement in Saccharum spp.

BACKGROUND: Nitrogen is an essential element for sugarcane growth and development and is generally applied in the form of urea often much more than at recommended rates, causing serious soil degradation, particularly soil acidification, as well as groundwater and air pollution. In spite of the importance of nitrogen for plant growth, fewer reports are available to understand the application and biological role of N2 fixing bacteria to improve N2 nutrition in the sugarcane plant. RESULTS: In this study, a total of 350 different bacterial strains were isolated from rhizospheric soil samples of the sugarcane plants. Out of these, 22 isolates were selected based on plant growth promotion traits, biocontrol, and nitrogenase activity. The presence and activity of the nifH gene and the ability of nitrogen-fixation proved that all 22 selected strains have the ability to fix nitrogen. These strains were used to perform 16S rRNA and rpoB genes for their identification. The resulted amplicons were sequenced and phylogenetic analysis was constructed. Among the screened strains for nitrogen fixation, CY5 (Bacillus megaterium) and CA1 (Bacillus mycoides) were the most prominent. These two strains were examined for functional diversity using Biolog phenotyping, which confirmed the consumption of diverse carbon and nitrogen sources and tolerance to low pH and osmotic stress. The inoculated bacterial strains colonized the sugarcane rhizosphere successfully and were mostly located in root and leaf. The expression of the nifH gene in both sugarcane varieties (GT11 and GXB9) inoculated with CY5 and CA1 was confirmed. The gene expression studies showed enhanced expression of genes of various enzymes such as catalase, phenylalanine-ammonia-lyase, superoxide dismutase, chitinase and glucanase in bacterial-inoculated sugarcane plants. CONCLUSION: The results showed that a substantial number of Bacillus isolates have N-fixation and biocontrol property against two sugarcane pathogens Sporisorium scitamineum and Ceratocystis paradoxa. The increased activity of genes controlling free radical metabolism may at least in part accounts for the increased tolerance to pathogens. Nitrogen-fixation was confirmed in sugarcane inoculated with B. megaterium and B. mycoides strains using N-balance and 15N2 isotope dilution in different plant parts of sugarcane. This is the first report of Bacillus mycoides as a nitrogen-fixing rhizobacterium in sugarcane.

Proteomic Analysis of the Resistance Mechanisms in Sugarcane during Sporisorium scitamineum Infection.

Smut disease is caused by Sporisorium scitamineum, an important sugarcane fungal pathogen causing an extensive loss in yield and sugar quality. The available literature suggests that there are two types of smut resistance mechanisms: external resistance by physical or chemical barriers and intrinsic internal resistance mechanisms operating at host⁻pathogen interaction at cellular and molecular levels. The nature of smut resistance mechanisms, however, remains largely unknown. The present study investigated the changes in proteome occurring in two sugarcane varieties with contrasting susceptibility to smut-F134 and NCo310-at whip development stage after S. scitamineum infection. Total proteins from pathogen inoculated and uninoculated (control) leaves were separated by two-dimensional gel electrophoresis (2D-PAGE). Protein identification was performed using BLASTp and tBLASTn against NCBI nonredundant protein databases and EST databases, respectively. A total of thirty proteins spots representing differentially expressed proteins (DEPs), 16 from F134 and 14 from NCo310, were identified and analyzed by MALDI-TOF/TOF MS. In F134, 4 DEPs were upregulated and nine were downregulated, while, nine were upregulated and three were downregulated in NCo310. The DEPs were associated with DNA binding, metabolic processes, defense, stress response, photorespiration, protein refolding, chloroplast, nucleus and plasma membrane. Finally, the expression of CAT, SOD, and PAL with recognized roles in S. scitamineum infection in both sugarcane verities were analyzed by real-time quantitative PCR (RT-qPCR) technique. Identification of genes critical for smut resistance in sugarcane will increase our knowledge of S. scitamineum-sugarcane interaction and help to develop molecular and conventional breeding strategies for variety improvement.

Magnetic Resonance Imaging (MRI): Operative Findings Correlation in 229 Fistula-in-Ano Patients.

BACKGROUND: To correlate the operative findings of patients with fistula-in-ano with preoperative MRI and quantify the information added with MRI. METHODS: All consecutive fistula-in-ano patients operated between July 2013 and May 2015 were prospectively enrolled. Preoperative MRI was done in every patient. The details of tracts, internal opening and "complex parameters" (additional tract or additional internal opening, horseshoe tract, associated abscess and supralevator extension) found at surgery were compared to the findings determined by MRI. RESULTS: A total of 229 patients (424 tracts) with mean age-49.0 ± 11.3 years were included. M/F 198/31. James hospital classification: Type I 58, II 20, III 49, IV 86 and V 16. The sensitivity and specificity of MRI in diagnosing fistula tracts were 98.8 and 99.7%, respectively, and in identifying internal opening were 97.7 and 98.6%, respectively. MRI added significant information in 46.7% (107/229) patients which was presence of additional tracts in 71 (66.3%), horseshoe tract in 63 (58.8%), supralevator extension in 16 (14.9%), unsuspected abscess in 11 (10.3%) and multiple internal openings in one patient (1%). The proportion of simple/complex fistula (based on history and clinical examination alone) was 32.8/67.2% which changed to 21.4/78.6% after the MRI scan. MRI added significant information about unsuspecting complex parameters which were missed on history and clinical examination in more than one-third (26/75: 34.6%) of simple fistulae and more than half (81/154: 52.5%) of already known complex fistulae. CONCLUSIONS: MRI is highly accurate in diagnosing fistula-in-ano and added significant information about unsuspected complex parameters in over one-third (34.6%) of simple and in half (52.5%) of complex fistula-in-ano.

Synergistic influence of Vetiveria zizanioides and selected rhizospheric microbial strains on remediation of endosulfan contaminated soil.

Application of endosulfan tolerant rhizospheric bacterial strain isolated from pesticide contaminated area, Ghaziabad in combination with V. zizanioides for the remediation of endosulfan is described herein. The dissipation of endosulfan from soil was considerably enhanced in the presence of bacterial strain and Vetiveria zizanioides together when compared to the dissipation in presence of either of them alone. Four strains- EAG-EC-12 (M1), EAG-EC-13(M2), EAG-EC-14(M3) and EAG-EC-15(M4) are used for this purpose. V. zizanioides was grown in garden soil spiked with 1500 µg g(-1) of endosulfan and inoculated with 100 ml of microbial culture of above motioned strains. Effect of microbial inoculation on plant growth, endosulfan uptake and endosulfan removal efficiency were analyzed. The microbial inoculation significantly enhances the growth of test plant and endosulfan dissipation from soil (p < 0.05). The addition of bacterial strain M1, M2, M3 and M4 in treated pots showed enhanced root length by 13, 33 35, 20.2 and 4.3 %, above ground plant length by 16.38, 35.56, 24.92 and 9.8 % and biomass by 33.69, 49.63, 39.24 and 17.09 % respectively when compared with endosulfan treated plants. After 135 days of exposure, a decline in endosulfan concentration by 59.12, 64.56, 62.69 and 56.39 % was obtained in the spiked soil inoculated with bacterial strains M1, M2, M3 and M4 respectively whereas, decrease in endosulfan concentration by 72.78, 85.25, 76.91 and 65.44 % in the vegetative spiked soil inoculated with same strains was observed during same exposure period. After 135 days of growth period, enhanced removal of endosulfan from experimental soil by 13.66, 20.69, 14.22 and 9.05 % was found in vegetative experiment inoculated with same strains when compared with non vegetative experiment. Result of the study showed that use of toletant plant and tolerant bacterial strains could be the better strategy for the remediation of endosulfan contaminated soil.

Design and synthesis of new fluconazole analogues.

We have synthesized new fluconazole analogues containing two different 1,2,3-triazole units in the side chain. The synthesis of new amide analogues using a variety of acids is also described. All the compounds showed very good antifungal activity. A hemolysis study of the most active compounds 6e and 13j showed that both compounds did not cause any hemolysis at the dilutions tested. These compounds did not exhibit any toxicity to L929 cells at MIC and lower concentrations. In the docking study, the overall binding mode of 6e and 13j appeared to be reasonable and provided a good insight into the structural basis of inhibition of Candida albicans Cyp51 by these compounds.

Synthesis and evaluation of new diaryl ether and quinoline hybrids as potential antiplasmodial and antimicrobial agents.

Synthesis and bioevaluation of new diaryl ether hybridized quinoline derivatives as antiplasmodial, antibacterial and antifungal agents is reported. It was encouraging to discover that several compounds displayed 2-3 folds better efficacy than chloroquine in chloroquine-resistant K1 strain of Plasmodium falciparum. Further, a few members of the library displayed good antibacterial efficacy against gram positive strains of bacteria but none of the compounds displayed any significant antifungal activity.

A unique dithiocarbamate chemistry during design & synthesis of novel sperm-immobilizing agents.

1-Substituted piperazinecarbodithioates were obtained by an unusual removal of CS2 in benzyl substituted dithiocarbamate derivatives under acid and basic conditions during design and synthesis of 1,4-(disubstituted)piperazinedicarbodithioates as double edged spermicides. A plausible mechanism for CS2 removal has been proposed. All synthesized compounds were subjected to spermicidal, antitrichomonal and antifungal activities. Twenty-one compounds irreversibly immobilized 100% sperm (MEC, 0.06-31.6 mM) while seven compounds exhibited multiple activities. Benzyl 4-(2-(piperidin-1-yl)ethyl) piperazine-1-(carbodithioate) (18) and 1-benzyl 4-(2-(piperidin-1-yl)ethyl)piperazine-1,4-bis(carbodithioate) (24) exhibited appreciable spermicidal (MEC, 0.07 and 0.06 mM), antifungal (MIC, 0.069-0.14 and >0.11 mM) and antitrichomonal (MIC, 1.38 and 0.14 mM) activities. The probable mode of action of these compounds seems to be through sulfhydryl binding which was confirmed by fluorescence labeling of sperm thiols.

Identification and characterization of ethanol utilizing fungal flora of oil refinery contaminated soil.

The indigenous fungal flora of three oil refinery contaminated sites (Bharuch, Valsad and Vadodara) of India has been documented in the present investigation. A total seventy-five fungal morphotypes were isolated from these sites and out of them, only fifteen isolates were capable of utilizing ethanol (0-8%; v:v) as a sole source of carbon and energy for growth. Ten percent ethanol was completely lethal for the growth of all the isolated fungus. Biochemical characterization of the potent ethanol utilizing fungal isolates was studied based on substrate utilization profiles using BIOLOG phenotype microarray plates. Based on the morphological characters and Internal Transcribed Spacer region of ribosomal DNA, the fungal isolates were identified as Fusarium brachygibbosum, Fusarium equiseti, Fusarium acuminatum, Pencillium citrinum, Alternaria tenuissima, Septogloeum mori, Hypocrea lixii, Aureobasidium sp., Penicillium sp., and Fusarium sp. Intra-species genetic diversity among Fusarium sp. was evaluated by whole genome analysis with repetitive DNA sequences (ERIC, REP and BOX) based DNA fingerprinting. It was found that these fungus use alcohol dehydrogenase and acetaldehyde dehydrogenase enzymes based metabolism pathway to utilize ethanol for their growth and colonization.

Optimization of media components for chitinase production by chickpea rhizosphere associated Lysinibacillus fusiformis B-CM18.

Chitinase producing strain B-CM18 was isolated from chickpea rhizosphere and identified as Lysinibacillus fusiformis B-CM18. It showed in vitro antifungal activity against a wide range of fungal plant pathogens and was found to produce several PGPR activities. Further, a multivariate response surface methodology was used to evaluate the effects of different factors on chitinolytic activity and optimizing enzyme production. A central composite design was employed to achieve the highest chitinase production at optimum values of the process variables, viz., temperature (20-45 °C), sodium chloride (2-7%), starch (0.1-1%) and yeast extract (0.1-1%), added in the minimal medium supplemented with colloidal chitin (1-10%; w:w). The fit of the model (R(2) = 0.5692) was found to be significant. The production medium to achieve the highest chitinase production (101 U ml(-1) ) was composed of the minimal medium composed of chitin (6.09%), NaCl (4.5%), starch (0.55%) and yeast extract (0.55%) with temperature (32.5 °C). The results show that the optimization strategy led to an increase in chitinase production by 56.1-fold. The molecular mass of the chitinase was estimated to be 20 kDa by anion exchange and gel filtration chromatography. Further, purified chitinase showed strong antifungal activity against test pathogens. Overall, these results may serve as a base line data for enhancing the chitinolytic potential of bacterial antagonists for bio-management of chickpea pathogens.

Pseudomonas aeruginosa Strain 91: A Multifaceted Biocontrol Agent against Banana Fusarium Wilt.

Banana Fusarium wilt (BFW), caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (Foc), poses significant threats to banana cultivation. Currently, effective control methods are lacking, and biological control has emerged as a possible strategy to manage BFW outbreaks. In this investigation, 109 bacterial strains were isolated from the rhizospheric soil surrounding banana plants in search of potent biological agents against Foc. Strain 91 exhibited the highest antifungal activity against the causal agent of Foc and was identified as Pseudomonas aeruginosa through 16S rRNA gene sequencing and scanning electron microscopy (SEM). Elucidation of strain 91's inhibitory mechanism against Foc revealed a multifaceted antagonistic approach, encompassing the production of bioactive compounds and the secretion of cell wall hydrolytic enzymes. Furthermore, strain 91 displayed various traits associated with promoting plant growth and showed adaptability to different carbon sources. By genetically tagging with constitutively expressing GFP signals, effective colonization of strain 91 was mainly demonstrated in root followed by leaf and stem tissues. Altogether, our study reveals the potential of P. aeruginosa 91 for biocontrol based on inhibition mechanism, adaptation, and colonization features, thus providing a promising candidate for the control of BFW.

Genome analysis of a halophilic Virgibacillus halodenitrificans ASH15 revealed salt adaptation, plant growth promotion, and isoprenoid biosynthetic machinery.

Globally, due to widespread dispersion, intraspecific diversity, and crucial ecological components of halophilic ecosystems, halophilic bacteria is considered one of the key models for ecological, adaptative, and biotechnological applications research in saline environments. With this aim, the present study was to enlighten the plant growth-promoting features and investigate the systematic genome of a halophilic bacteria, Virgibacillus halodenitrificans ASH15, through single-molecule real-time (SMRT) sequencing technology. Results showed that strain ASH15 could survive in high salinity up to 25% (w/v) NaCl concentration and express plant growth-promoting traits such as nitrogen fixation, plant growth hormones, and hydrolytic enzymes, which sustain salt stress. The results of pot experiment revealed that strain ASH15 significantly enhanced sugarcane plant growth (root shoot length and weight) under salt stress conditions. Moreover, the sequencing analysis of the strain ASH15 genome exhibited that this strain contained a circular chromosome of 3,832,903 bp with an average G+C content of 37.54%: 3721 predicted protein-coding sequences (CDSs), 24 rRNA genes, and 62 tRNA genes. Genome analysis revealed that the genes related to the synthesis and transport of compatible solutes (glycine, betaine, ectoine, hydroxyectoine, and glutamate) confirm salt stress as well as heavy metal resistance. Furthermore, functional annotation showed that the strain ASH15 encodes genes for root colonization, biofilm formation, phytohormone IAA production, nitrogen fixation, phosphate metabolism, and siderophore production, which are beneficial for plant growth promotion. Strain ASH15 also has a gene resistance to antibiotics and pathogens. In addition, analysis also revealed that the genome strain ASH15 has insertion sequences and CRISPRs, which suggest its ability to acquire new genes through horizontal gene transfer and acquire immunity to the attack of viruses. This work provides knowledge of the mechanism through which V. halodenitrificans ASH15 tolerates salt stress. Deep genome analysis, identified MVA pathway involved in biosynthesis of isoprenoids, more precisely "Squalene." Squalene has various applications, such as an antioxidant, anti-cancer agent, anti-aging agent, hemopreventive agent, anti-bacterial agent, adjuvant for vaccines and drug carriers, and detoxifier. Our findings indicated that strain ASH15 has enormous potential in industries such as in agriculture, pharmaceuticals, cosmetics, and food.

Nitrogen fixation and phytohormone stimulation of sugarcane plant through plant growth promoting diazotrophic Pseudomonas.

Diazotrophic microorganisms are free-living groups of organisms that can convert atmospheric nitrogen (N) into bioavailable nitrogen for plants, which increases crop development and production. The purpose of the current study was to ascertain how diazotrophic plant growth promoting (PGP) Pseudomonas strains (P. koreensis CY4 and P. entomophila CN11) enhanced nitrogen fixation, defense activity, and PGP attributes of sugarcane varieties; GT11 and G×B9. A 15N isotope-dilution study was conducted to confirm the sugarcane strains' capacity to fix nitrogen, and the results indicated that between 21 to 35% of plant, nitrogen is fixed biologically by selected rhizobacteria. In comparison to the control, after 30, 60, and 90 days, both CY4 and CN11 strains significantly increased defense-related enzymes (catalase, peroxidase, phenylalanine ammonia-lyase, superoxide dismutase, glucanase, and chitinase) and phytohormones (abscisic acid, ABA, cytokinin, etc.) in GT11 and GXB. Additionally, the expression of SuCHI, SuGLU, SuCAT, SuSOD, and SuPAL genes was found to be elevated in Pseudomonas strains inoculated plants using real-time quantitative polymerase chain reaction (RT-qPCR). Both bacterial strains increased all physiological parameters and chlorophyll content in sugarcane plants more than their control. The effects of P. koreensis CY4 and P. entomophila CN11 strains on sugarcane growth promotion and nitrogen fixation under greenhouse conditions are described here for the first time systematically. The results of confirmation studies demonstrated that P. koreensis CY4 and P. entomophila are PGP bacterial strains with the potential to be employed as a biofertilizer for sugarcane growth, nitrogen nutrient absorption, and reduced application of chemical nitrogenous fertilizers in agricultural fields. .

Effect of endophytic diazotroph Enterobacter roggenkampii ED5 on nitrogen-metabolism-related microecology in the sugarcane rhizosphere at different nitrogen levels.

Sugarcane is an important sugar and energy crop worldwide, requiring a large amount of nitrogen (N). However, excessive application of synthetic N fertilizer causes environmental pollution in farmland. Endophytic nitrogen-fixing bacteria (ENFB) provide N nutrition for plants through biological N fixation, thus reducing the need for chemical fertilizers. The present study investigated the effect of the N-fixing endophytic strain Enterobacter roggenkampii ED5 on phytohormone indole-3-acetic acid (IAA), N-metabolism enzyme activities, microbial community compositions, and N cycle genes in sugarcane rhizosphere soil at different N levels. Three levels of 15N-urea, such as low N (0 kg/ha), medium N (150 kg/ha), and high N (300 kg/ha), were applied. The results showed that, after inoculating strain ED5, the IAA content in sugarcane leaves was significantly increased by 68.82% under low N condition at the seedling stage (60 days). The nitrate reductase (NR) activity showed a downward trend. However, the glutamine synthase (GS) and NADH-glutamate dehydrogenase (NADH-GDH) activities were significantly enhanced compared to the control under the high N condition, and the GS and NR genes had the highest expression at 180 and 120 days, respectively, at the low N level. The total N content in the roots, stems, and leaves of sugarcane was higher than the control. The 15N atom % excess of sugarcane decreased significantly under medium N condition, indicating that the medium N level was conducive to N fixation in strain ED5. Metagenome analysis of sugarcane rhizosphere soil exhibited that the abundance of N-metabolizing microbial richness was increased under low and high N conditions after inoculation of strain ED5 at the genus level, while it was increased at the phylum level only under the low N condition. The LefSe (LDA > 2, p < 0.05) found that the N-metabolism-related differential microorganisms under the high N condition were higher than those under medium and low N conditions. It was also shown that the abundance of nifDHK genes was significantly increased after inoculation of ED5 at the medium N level, and other N cycle genes had high abundance at the high N level after inoculation of strain ED5. The results of this study provided a scientific reference for N fertilization in actual sugarcane production.