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Abstract In the Kandi zone of Punjab, India, root and rhizospheric soil samples were collected from the local vegetation near the Shivalik mountain foothills. Fifteen fungal colonies exhibiting distinct cultural morphology on Potato Dextrose Agar (PDA) plates were selected for plant–microbe interaction studies. Among these, the isolate HNB9 was identified as a nonpathogenic root colonizer. Morphological and molecular analyses confirmed HNB9 as Talaromyces albobiverticillius, characterized by the secretion of a red pigment as a secondary metabolite. Plants colonized with T. albobiverticillius HNB9 exhibited enhanced growth, manifesting in increased shoot and root length compared to untreated controls. This study unveiled the first evidence that a species from the Talaromyces genus, specifically T. albobiverticillius, possesses dual capabilities of root colonization and plant growth promotion. Moreover, HNB9 demonstrated the production of plant growth-regulating compounds like Indole Acetic Acid (IAA) and proficient solubilization of crucial nutrients (Phosphorous, Zinc, and Silica) through plate culture methods. This finding represents a significant contribution to the understanding of root-colonizing fungi with plant growth-promoting attributes, challenging the existing knowledge gap within the Talaromyces genus.

Biotechnological and industrial processes involve applications of various microorganisms and enzymes, and laccase, as a multifunctional enzyme, is admired for its role in degrading a variety of substances. Laccase is a copper-containing oxidase enzyme that is usually found in insects, plants, and microorganisms including fungi and archaea. Several phenolic substrates are oxidized by laccases, which results in crosslinking. Various research work and industrial solutions have identified the true potential of laccases to degrade various aromatic polymers, and their plausible application in bioremediation and other industries is entirely conceivable. This review focuses on laccases as a multifunctional enzyme and provides an overview of its natural origin, catalytic mechanism, and various methods of production. Further, we discuss the various applications of laccase in the biotechnological arena. We observed that laccase can degrade and detoxify various synthetic compounds. The broad substrate specificity of the same makes it worthy for different fields of industrial applications such as food and bioremediation technology, textile and paper technology, biosensors and nanobiotechnology, biofuel, and various other applications, which are described in this paper. These recent developments in the application of laccase show the multifunctional role of laccase in industrial biotechnology and provide an outlook of laccase as a multifunctional enzyme at the forefront of biotechnology.

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.

Rapid urbanization and globalization demand increasing agricultural productivity. Soil nutrient supply capacity is continuously decreasing due to soil erosion, degradation, salt deposition, undesired element, metal deposition, water scarcity, and an uneven nutrient delivery system. Rice cultivation requires a large amount of water which is becoming detrimental due to these activities. There is a need to increase its productivity. Microbial inoculants are becoming increasingly important in achieving sustainable agricultural production systems. The current study was conducted to investigate the interaction between the root endophytic fungus Serendipita indica (S. indica) and the actinobacterium Zhihengliuella sp. ISTPL4 (Z. sp. ISTPL4) and their synergistic effects on the growth of rice (Oryza sativa L). Both S. indica and Z. sp. ISTPL4 showed positive interactions. Growth of S. indica was observed at different days after Z. sp. ISTPL4 inoculation, and stimulated growth of S. indica was observed when Z. sp. ISTPL4 was inoculated at 5 dafi (days after fungal inoculation). Z. sp. ISTPL4 promoted the growth of S. indica as it increased spore germination. Furthermore, confocal and scanning electron microscopy (SEM) analyses showed a 27% increase in the spore size of S. indica in the presence of Z. sp. ISTPL4. In a liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis increased production of alanine and glutamic acid was observed in their sequential co-culture as compared with individual cultures. Sequential inoculation of S. indica and Z. sp. ISTPL4 significantly increased the biochemical and physical characteristics of rice as compared with their individual inoculum. Biochemical parameters such as chlorophyll content, total soluble sugar, and flavonoid content in the rice increased by up to 57%, 47%, and 39%, respectively, in the presence of the combined inoculum of S. indica and Z. sp. ISTPL4. This will be the first study, to the best of our knowledge, which shows the fungus and actinobacterium interaction and their synergistic roles in the growth promotion of rice. Furthermore, this novel combination can also be used to boost the growth of other crops to increase the agricultural yield.

Halotolerant plant growth promoting rhizobacteria (PGPR) are beneficial microorganisms utilized to mitigate the biotic and abiotic stresses in plants. The areas of Sundarban mangroves of West Bengal, India have been reported to be rich in halotolerant microflora, yet major area remains unexplored. The present study, therefore, aims to map down the region-specific native microbial community potent of salt tolerance, plant growth promoting (PGP) activity and antagonistic activity against fungal pathogens. Bacterial samples were isolated from the saline soil of the Sundarban mangroves. A total of 156 bacterial samples were isolated and 20 were screened for their salt tolerance potential. These isolates were characterised using morphological, biochemical, and molecular approaches. Based on 16s rRNA sequencing, they were classified into 4 different genera, including Arthrobacter sp. (01 isolate), Pseudomonas plecoglossicida (01 isolate), Kocuria rosea (01 isolate), and Bacillus (17 isolates). The halotolerant isolates which possessed plant growth promoting traits including phosphate, and zinc solubilization, indole acetic acid production, siderophore, and ammonia generation were selected. Further, the effect of two halotolerant isolates GN-5 and JR-12 which showed most prominent PGP activities was evaluated in pea plant under high salinity conditions. The isolates improved survival by promoting germination (36 to 43%) and root-shoot growth and weight of pea plant in comparison to non-inoculated control plants. In a subsequent dual culture confrontation experiment, both these halo-tolerant isolates showed antagonistic activities against the aggressive root rot disease-causing Macrophomina phaseolina (Tassi) Goid NAIMCC-F-02902. The identified isolates could be used as potential bioagents for saline soils, with potential antagonistic effect on root rot disease. However, further studies at the physiological and molecular level would help to delineate a detail mechanistic understanding of broad-spectrum defence against salinity and potential biotic pathogen.

Abstract Background Staphylococcus aureus is a notorious human pathogen that causes often lethal systemic conditions that are mostly medical device associated biofilm infections. Similarly, coagulase negative staphylococci are emerging as leading pathogen for nosocomial infections owing to their ability to form biofilm on implanted medical equipment. Chronic in nature, these infections are difficult to treat. Such recalcitrance of these infections is caused mainly due to the presence of persister cells, which exhibit transient yet extreme tolerance to antibiotics. Despite tremendous clinical significance, there is lack of studies on persister cells formation among clinical bacterial isolates. Considering the importance of factors influencing persister formation, in this study, we evaluate the association of antibiotic tolerance with biofilm production, antibiotic stress, growth phase, specimen type, and dependency on staphylococcal species. Biofilm formation was detected among 375 clinical staphylococcal isolates by quantitative tissue culture plate method (TCP) and icaAD genes by genotypic method. The antibiotic susceptibility was determined by Kirby Bauer disc diffusion method while minimum inhibitory concentration values were obtained by agar dilution method. Persister cells were measured in the susceptible staphylococcal isolates in the presence of clinically relevant antibiotics. Results In the study, 161 (43%) S. aureus and 214 (57%) coagulase negative staphylococci (CNS) were isolated from different clinical samples. TCP method detected biofilm production in 84 (52.2%) S. aureus and 90 (42.1%) CNS isolates. The genotypic method detected icaAD genes in 86 (22.9%) isolates. Majority (> 90%) of both the biofilm producers and non-producers were sensitive to chloramphenicol and tetracycline but were resistant to penicillin. Interestingly, all isolates were sensitive to vancomycin irrespective of biofilm production. While high persister frequency was observed among all staphylococci isolates in the stationary growth phase, the persister frequency in exponential growth phase was statistically high among isolates possessing icaAD genes compared to icaAD negative isolates. Conclusion The research findings provide strong evidence that the clinical staphylococcal isolates exhibit extreme antibiotic tolerance suggesting their causal link with treatment failures. Understanding the factors influencing the formation and maintenance of persister cells are of utmost important aspect to design therapeutics and control recalcitrant bacterial infections.

This study investigates the mechanisms underlying the enhanced zinc (Zn) and iron (Fe) uptake in a low Zn- and Fe-accumulating Indian wheat genotype HD2967 upon co-inoculation of Piriformospora indica, a phyto-promotional fungal endophyte, and Azotobacter chroococcum, a rhizobacteria, in a hydroponic system. Modifications in root architecture, low molecular-weight organic acid (LMWOA) profile in root exudates, and expression of TaZIP genes in roots and shoots of treated and control plants were analysed. The LMWOA profiles of treated and control plants were investigated using high-performance liquid chromatography, which showed the presence of four LMWOAs: citric, malic, acetic, and succinic acids. A 3.6-fold increase was observed in the total organic acid content of colonised over control plants with citric acid being predominant – representing 90% of the total organic acids. Microbial colonisation caused profuse branching in roots of treated plants with increases in root volume, total surface area, total length, and numbers of tips, forks, and crossings by 1.39-, 1.37-, 1.67-, 1.4-, 1.32-, and 1.81-fold, respectively. Four targeted TaZIP genes (TaZIP1, TaZIP3, TaZIP5, and TaZIP7) were overexpressed in roots and shoots of treated plants as compared to control plants. The results clearly suggest that endophytic colonisation modified the LMWOA profile in root exudates, root morphology, and expression of TaZIP genes in roots and shoots, of colonised hosts would have synergistically facilitated enhanced accumulation of Fe and Zn in colonised plants.

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.

The biological synthesis of nanoparticles using fungal cultures is a promising and novel tool in nano-biotechnology. The potential culture of Trichoderma asperellum (T. asperellum) has been used to synthesize copper oxide nanoparticles (CuO NPs) in the current study. The necrotrophic infection in Brassica species is caused due to a foliar pathogen Alternaria brassicae (A. brassicae). Mycogenic copper oxide nanoparticles (M-CuO NPs) were characterized by spectroscopic and microscopic techniques such as UV–visible spectrophotometry (UV–vis), transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The antifungal potential of CuO NPs was studied against A. brassicae. M-CuO NPs exhibited a surface plasmon resonance (SPR) at 303 nm, and XRD confirmed the crystalline phase of NPs. FTIR spectra confirmed the stretching of amide bonds, and the carbonyl bond indicated the presence of enzymes in T. asperellum filtrate. SEM and TEM confirmed the spherical shape of M-CuO NPs with an average size of 22 nm. Significant antifungal potential of M-CuO NPs was recorded, as it inhibited the growth of A. brassicae up to 92.9% and 80.3% in supplemented media with C-CuO NPs at 200 ppm dose. Mancozeb and propiconazole inhibited the radial growth up to 38.7% and 44.2%. SEM confirmed the morphological changes in hyphae and affected the sporulation pattern. TEM revealed hardly recognizable organelles, abnormal cytoplasmic distribution, and increased vacuolization, and light microscopy confirmed the conidia with reduced diameter and fewer septa after treatment with both types of NPs. Thus, M-CuO NPs served as a promising alternative to fungicides.

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.

Abstract Background Coagulase-negative staphylococci (CNS) survive as commensals of skin, anterior nares and external canals of human and were regarded as non-infectious pathogens. However, they are emerging as a major cause of nosocomial infectious due to their ability to form biofilms and high resistance to several classes of antibiotics. This study examines the biofilm forming abilities of 214 clinical CNS isolates using phenotypic and genotypic methods, and determines their antibiotic susceptibility patterns. Methods A total of 214 clinical isolates collected from different clinical samples were identified as CNS and their antibiotic susceptibility determined by CLSI guidelines. The biofilm forming ability of all isolates was determined by three phenotypic methods; Congo red agar (CRA) method, tube adherence method (TM) and tissue culture plate (TCP) method and by genotypic method for the detection of icaAD genes. Results Among all the isolates, S. epidermidis (57.5%) was found the most frequently, followed by S. saprophyticus (18.7%), S. haemolyticus (11.2%), S. hominis (7%), and S. capitis (5.6%). Antibiotic susceptibility pattern demonstrated 91.6% isolates were resistant to penicillin and 66.8% to cefoxitin while 91.1% isolates were susceptible to chloramphenicol. Constitutive and inducible clindamycin resistant phenotype as measured by D-test was seen among 28% and 14.5% of isolates respectively. Tissue culture plate method detected biofilm production in 42.1% isolate followed by 31.8% through tube method while 20.1% isolates were found to produce slime in Congo red agar method. The genotypic assay revealed presence of icaA and icaD genes in 19.2% isolates. Conclusion The study shows a high prevalence of biofilm formation and inducible clindamycin resistance in CNS isolates, indicating the importance of in-vitro biofilm production test and D-test in routine laboratory diagnostics. Implementation of efficient diagnostic techniques for detection of biofilm production in clinical samples can help manage staphylococcal infections and minimize risks of treatment failures in hospitals.

The use of biofertilizers has been the spotlight of research aiming to mitigate the food security threat as well as to restore the fertility of agricultural lands, for decades. Several studies are being conducted to unravel the role and mechanisms of plant growth-promoting microbes. In the present research, we evaluated the effect of silver nanoparticles (AgNPs) and Piriformospora indica on the growth and nutritional enhancement of black rice (Oryzae sativa. L.) individually and in combination. Among the different treatment conditions, the AgNPs + P. indica treatment led to a significant (p ≤ 0.05) increase in morphological and agronomic parameters. In comparison to the control, the percentage increase in plant height in AgNPs-treated black rice was 2.47%, while that for the treatment with only P. indica was 13.2% and that for the treatment with both AgNPs + P. indica was 30.9%. For the number of productive tillers, the effect of AgNPs in comparison to the control was non-significant; however, the effect of P. indica and AgNPs + P. indica showed a significant (p ≤ 0.05) increase of 13.2% and 30.9% in both the treatments, respectively. Gas chromatography mass spectrophotometry analysis of grains revealed that the contents of phenylalanine, tryptophan, and histidine (aromatic amino acids) were significantly (p ≤ 0.05) increased by 75%, 11.1%, and 50%, respectively, in P. indica-treated black rice. Nutrient profiling showed that macronutrients such as potassium, calcium, magnesium were found to be increased by 72.8%, 86.4% and 59.2%, respectively, in the treatment with AgNPs + P. indica in comparison to the control plants. Additionally, a significant (p ≤ 0.05) increase of 51.9% in anthocyanin content was observed in AgNPs + P. indica-treated black rice. The P. indica treatment also showed improved growth and augmented nutrient contents. From this study, we were able to understand that AgNPs + P. indica treatment would be a better plant growth-promoting factor and further evaluation would enable us to obtain a clear picture of its mechanisms of action.

Ruminant-origin food products are the richest source of natural conjugated linoleic acid (CLA) in human diets, and cis-9, trans-11 (c9, t11) is the most abundant isomer of CLA. Butyrivibrio fibrisolvens, a rumen microbe, is considered responsible to have the highest CLA-producing potential. The present study was carried out to identify and characterize a rumen-origin B. fibrisolvens strain that can produce high levels of c9, t11 CLA, as well as to evaluate how dietary supplementation with the strain influenced c9, t11 CLA levels in the gut and adipose tissue in mice. A total of 257 anaerobic strains were isolated from buffalo, cattle, and goat rumen samples, and 24 isolates were identified as Butyrivibrio spp. based on 16S rRNA sequencing. 22 isolates of Butyrivibrio spp. were found to be B. fibrisolven-like. These strains were tested for their potential to produce CLA at varied concentrations (0, 100, 200, 300, 400, 500 μg/mL) of linoleic acid (LA) as substrate after incubating for different durations (0, 2, 4, 6, 12, 24 and 36 h). Results of the study reflected that Butyrivibrio fibrisolvens F7 converted almost 74% of LA of 200 μg/mL concentration to c9, t11 CLA after 24 h and formed the maximum CLA (147.39 μg/mL). B. fibrisolvens F7 (GenBank Accession No: KU507371) was found to be acid and bile tolerant and exhibited good cell surface hydrophobicity in n-hexadecane and toluene. B. fibrisolvens F7 dietary supplementation in mice, combined with a high-LA diet, increases c9, t11 CLA levels in adipose tissue and gut by more than three folds. The results of this study significantly increase the possibility of employing B. fibrisolvens F7 as a probiotic supplement for animals to maintain a steady supply of natural c9, t11 CLA in the host.

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.

Synthetic dyes pose a large threat to the environment and consequently to human health. Various dyes are used in textile, cosmetics, and pharmaceutical industries, and are released into the environment without any treatment, thus adversely affecting both the environment and neighboring human populations. Several existing physical and chemical methods for dye degradation are effective but have many drawbacks. Biological methods over the years have gained importance in the decolorization and degradation of dye and have also overcome the disadvantages of physiochemical methods. Furthermore, biological methods are eco-friendly and lead to complete decolorization. The mechanism of decolorization and degradation by several bacterial enzymes are discussed in detail. For the identification of ecologically sustainable strains and their application at the field level, we have focused on bioaugmentation aspects. Furthermore, in silico studies such as molecular docking of bacterial enzymes with dyes can give a new insight into biological studies and provide an easy way to understand the interaction at the molecular level. This review mainly focuses on an integrative approach and its importance for the effective treatment and decolorization of dyes.

Synthetic dyes and colourants have been the mainstay of the pigment industry for decades. Researchers are eager to find a more environment friendly and non-toxic substitute because these synthetic dyes have a negative impact on the environment and people’s health. Microbial pigments might be an alternative to synthetic pigments. Microbial pigments are categorized as secondary metabolites and are mainly produced due to impaired metabolism under stressful conditions. These pigments have vibrant shades and possess nutritional and therapeutic properties compared to synthetic pigment. Microbial pigments are now widely used within the pharmaceuticals, food, paints, and textile industries. The pharmaceutical industries currently use bacterial pigments as a medicine alternative for cancer and many other bacterial infections. Their growing popularity is a result of their low cost, biodegradable, non-carcinogenic, and environmentally beneficial attributes. This audit article has made an effort to take an in-depth look into the existing uses of bacterial pigments in the food and pharmaceutical industries and project their potential future applications.

In the current research, unique Nano-Embedded Fungus (NEF), made by the synergic association of silver nanoparticles (AgNPs) and endophytic fungus (Piriformospora indica), is studied, and the impact of NEF on black rice secondary metabolites is reported. AgNPs were synthesized by chemical reduction process using the temperature-dependent method and characterized for morphological and structural features through UV visible absorption spectroscopy, zeta potential, XRD, SEM-EDX, and FTIR spectroscopy. The NEF, prepared by optimizing the AgNPs concentration (300 ppm) in agar and broth media, showed better fungal biomass, colony diameter, spore count, and spore size than the control P. indica. Treatment with AgNPs, P. indica, and NEF resulted in growth enhancement in black rice. NEF and AgNPs stimulated the production of secondary metabolites in its leaves. The concentrations of chlorophyll, carotenoids, flavonoids, and terpenoids were increased in plants inoculated with P. indica and AgNPs. The findings of the study highlight the synergistic effect of AgNPs and the fungal symbionts in augmenting the secondary metabolites in leaves of black rice.

Background: Microbial community analysis of electronic waste (e-waste)-polluted environments is of interest to understand the effect of toxic e-waste pollutants on the soil microbial community and to evaluate novel microorganisms resisting the toxic environment. The present study aims to investigate the bacterial community structure in soils contaminated with e-waste from various sites of Loni and Mandoli (National Capital Region (NCR), India) where e-waste dumping and recycling activities are being carried out for many years. Results: Interferences to soil metagenomic DNA extraction and PCR amplification were observed because of the presence of inhibiting components derived from circuit boards. Whole-metagenome sequencing on the Illumina MiSeq platform showed that the most abundant phyla were Proteobacteria and Firmicutes. Deltaproteobacteria and Betaproteobacteria were the most common classes under Proteobacteria. Denaturing gradient gel electrophoresis (DGGE) analysis of the bacterial 16S rRNA gene showed that e-waste contamination altered the soil bacterial composition and diversity. There was a decrease in the number of predominant bacterial groups like Proteobacteria and Firmicutes but emergence of Actinobacteria in the contaminated soil samples. Conclusions: This is the first report describing the bacterial community structure of composite soil samples of e-waste-contaminated sites of Loni and Mandoli, Delhi NCR, India. The findings indicate that novel bacteria with potential bioremediating properties may be present in the e-waste-contaminated sites and hence need to be evaluated further.How to cite: Salam M, Varma A. Bacterial community structure in soils contaminated with electronic waste pollutants from Delhi NCR, India. Electron J Biotechnol 2019;41. https://doi.org/10.1016/j.ejbt.2019.07.003. Keywords: Bacterial community, Contaminated soils, DGGE, Electronic waste, e-Waste, Heavy metals, Humic substances, India, Metagenomics, Organic pollutants, Proteobacteria

Abstract Objective Geographic expansion of dengue incidence has drawn a global interest to identify the influential factors that instigate the spread of this disease. The objective of this study was to find the environmental factors linked to dengue incidence in a dengue epidemic area of Nepal by negative binomial models using climatic factors from 2010 to 2017. Results Minimum temperature at lag 2 months, maximum temperature and relative humidity without lag period significantly affected dengue incidence. Rainfall was not associated with dengue incidence in Chitwan district of Nepal. The incident rate ratio (IRR) of dengue case rise by more than 1% for every unit increase in minimum temperature at lag 2 months, maximum temperature and relative humidity, but decrease by .759% for maximum temperature at lag 3 months. Considering the effect of minimum temperature of previous months on dengue incidence, the vector control and dengue management program should be implemented at least 2 months ahead of dengue outbreak season.

Abstract Background The expansion of dengue vectors from lowland plains to the upland hilly regions of Nepal suggests the likelihood of increased risk of dengue. Our objective was to assess the effects of meteorological variables on vector indices and populations of dengue vectors in two different ecological regions of Nepal. An entomological survey was conducted in Kathmandu and Lalitpur (upland) and Chitwan (lowland) of Nepal in three different seasons from July 2015 to May 2016. The effect of meteorological variables on vector indices (house index, container index and Breteau index) and Aedes spp. population abundance was analyzed. A gamma regression was used to fit the models for vector indices and a negative binomial regression was used to model Aedes spp. population abundance. Results Monsoon season showed higher values for vector indices and vector populations compared to post-monsoon and pre-monsoon. Overall, the factor temperature-rainfall effect had a more significant influence on vector indices compared to relative humidity. The regression models showed that relative humidity has a greater impact in Chitwan than in Kathmandu. Variation was observed in the effect of predictor variables on Aedes aegypti and Ae. albopictus abundance. Conclusions Temperature and rainfall contribute to the vector indices in the upland hilly region while relative humidity contributes in the lowland plains. Since vector prevalence is not only linked to meteorological factors, other factors such as water storage practices, waste disposal, sanitary conditions and vector control strategy should also be considered. We recommend strengthening and scaling up dengue vector surveillance and control programmes for monsoon season in both upland and lowland regions in Nepal.

The present scenario of agricultural sector is dependent hugely on the use of chemical-based fertilizers and pesticides that impact the nutritional quality, health status, and productivity of the crops. Moreover, continuous release of these chemical inputs causes toxic compounds such as metals to accumulate in the soil and move to the plants with prolonged exposure, which ultimately impact the human health. Hence, it becomes necessary to bring out the alternatives to chemical pesticides/fertilizers for improvement of agricultural outputs. The rhizosphere of plant is an important niche with abundant microorganisms residing in it. They possess the properties of plant growth promotion, disease suppression, removal of toxic compounds, and assimilating nutrients to plants. Utilizing such beneficial microbes for crop productivity presents an efficient way to modulate the crop yield and productivity by maintaining healthy status and quality of the plants through bioformulations. To understand these microbial formulation compositions, it becomes essential to understand the processes going on in the rhizosphere as well as their concrete identification for better utilization of the microbial diversity such as plant growth–promoting bacteria and arbuscular mycorrhizal fungi. Hence, with this background, the present review article highlights the plant microbiome aboveground and belowground, importance of microbial inoculants in various plant species, and their subsequent interactive mechanisms for sustainable agriculture.

Fusarium wilt caused by Fusarium oxysporum f. sp. lentis (Fol) has been considered as destructive seed and soil-borne disease of lentil which causes severe yield loss every year. For its biological control, twenty isolates of Trichoderma spp. were tested against thirty Fol isolates using dual culture technique. Out of twenty Trichoderma isolates, seven isolates (5593, ThL-1, ThL-4, TaL- 2, TaL- 4, ThaL-3 and TvL-6) showed maximum percent inhibition of radial growth against pathogens between 82.4% to 78.0% as compared to the rest of Trichoderma isolates. As far as molecular identification is concerned the effective six isolates after dual culture assay and highly virulent Fol isolate were molecularly identified by using ITS primers (ITS 1 and ITS 4) and remaining one isolate was taken from ITCC which was already identified as T. harzianum (5593). Seven efficient isolates of Trichoderma were used to evaluate volatile and non- volatile assay against Fol (NFLRJ-3). Isolates, 5593 and ThL-4 showed significant growth inhibition against Fol at 48 hours of incubation in both the assays as compared to the rest Trichoderma isolates. In volatile organic compound (VOC) assay ThL-4 showed highest number of seed germination and showed good seed health condition. Out of seven selected isolates of Trichoderma, all isolates were found effective to control disease and stimulated plant growth promotion activity of lentil (2017-2019). Therefore, these isolates can be utilized to formulate bio-fungicides which is the need of the hour as far as disease management is concerned.

Serendipita indica (former Piriformospora indica) is a non-obligate endophytic fungus and generally a plant growth and defence promoter with high potential to be used in agriculture. However, S. indica may switch from biotrophy to saprotrophy losing its plant growth promoting traits. Our aim was to understand if the free-living stage growth conditions (namely C availability) regulate S. indica’s phenotype, and its potential as plant-growth-promoting-microbe (PGPM). We grew S. indica in its free-living stage under increasing C availabilities (2–20 g L–1 of glucose or sucrose). We first characterised the effect of C availability during free-living stage growth on fungal phenotype: colonies growth and physiology (plasma membrane proton pumps, stable isotopic signatures, and potential extracellular decomposing enzymes). The effect of the C availability during the free-living stage of the PGPM was evaluated on wheat. We observed that C availability during the free-living stage regulated S. indica’s growth, ultrastructure and physiology, resulting in two distinct colony phenotypes: compact and explorer. The compact phenotype developed at low C, used peptone as the major C and N source, and displayed higher decomposing potential for C providing substrates; while the explorer phenotype developed at high C, used glucose and sucrose as major C sources and casein and yeast extract as major N sources, and displayed higher decomposing potential for N and P providing substrates. The C availability, or the C/N ratio, during the free-living stage left a legacy to the symbiosis stage, regulating S. indica’s potential to promote plant growth: wheat growth promotion by the explorer phenotype was ± 40% higher than that by the compact phenotype. Our study highlights the importance of considering microbial ecology in designing PGPM/biofertilizers. Further studies are needed to test the phenotypes under more extreme conditions, and to understand if the in vitro acquired characteristics persist under field conditions.

Abstract Objective Staphylococcus genus comprising both Staphylococcus aureus and coagulase negative staphylococci (CoNS) are widely distributed in nature and can infect diversity of hosts. Indeed, staphylococci are the major pathogens causing biofilm associated infections caused by contaminated hospital indwelling devices. These infections are persistent in nature being highly refractory to various stresses including antibiotics. Implementation of efficient diagnostic techniques for the biofilm production would help minimize the disease burden. Thus, early detection of pathogenic strains producing biofilms warrant the utmost importance in diagnostic laboratories especially in resource limited settings. Result Among 375 isolates collected from different clinical specimens, 214 (57%) were identified as coagulase negative staphylococci and 161 (43%) S. aureus. Detection of In-vitro biofilm formation in these isolates were carried out by three commonly used phenotypic assays and a genotypic assay. While evaluating the results, tissue-culture method with supplemented glucose and sucrose showed the best correlation with the results of genotypic assay.

Piriformospora indica is a root endophytic fungus that build a symbiotic association with the roots of a vascular host plants and induces overall plant growth. The P.indica can grown on a variety of synthetic and complex media. But these readymade medium consist of about 20-25 expensive chemicals. To minimise the cost for P.indica production (Arora et al 2014 and Varma et al, 2015) have already standardized a cost effective Jaggery medium (4% w/v) as a energy nutrient source for mass cultivation of P. indica with no compromise on quality. But growth of fungus varies substantially with different sources of Jaggery. So we collect Jaggery from four different sources i.e from 1. Local shop, 2. Indian Institute of Sugarcane Research, Lucknow, 3. Safal, New Delhi and 4.Organic Jaggery to check its impact on fungal growth and also check its efficacy on Spinach (Spinacia oleracea). The data observed from the present study shows that maximum growth of P.indica in terms of dry cell weight, radial diameter and spore count was recorded on Safal Jaggery medium as compared to other Jaggery medium. Moreover, fungal biomass grown on this medium showed positive effect on growth of spinach as it leads to increase in plant height and its dry and fresh weight. This finding suggest that Safal Jaggery medium is a cheaper and best source for commercial cultivation of P.indica.

The subsequent application of insecticides and pesticides in agriculture results in several health and environment issues. To overcome, such devastating effect of synthetic chemicals, an eco friendly measure is required. Chitinolytic bacteria and their enzymes can be adopted as potent substitutes to chemicals required controlling the agriculture loss. The aim of this research was to isolate bacterial strains with significant chitinase activity. Isolates were screened based on method viz. zone inhibition, colorimetric, biochemical and were identified based on 16S rDNA sequences. Observed chitinase activity was in range 0.181Uml-1 to 1.594Uml-1 with zone inhibition in the range of 6mm to 29mm. Among the recovered strains only two, MSCP10 and MSCW8 showed good response when tested against insect and showed 80% and 95% mortality respectively after 72 hours of treatment. Based on 16S rDNA sequencing, MSCP10 and MSCW8 exhibited similarity with Serratia marcescens strain S308 and Staphylococcaceae bacterium HDMd_5 respectively. Through insect bioassay it was concluded that these bacterial strains were effective against Lepidopteran insect P. xylostella.

Achieving food security in a context of environmental sustainability is one of the main challenges of the XXI century. Two competing strategies to achieve this goal are the use of genetically modified plants and the use of plant growth promoting microorganisms (PGPMs). However, few studies assess the response of genetically modified plants to PGPMs. The aim of this study was to compare the response of over-expressing the vacuolar H+-PPase (AVP) and wild-type rice types to the endophytic fungus; Piriformospora indica. Oryza sativa plants (WT and AVP) were inoculated with P. indica and 30 days later, morphological, ecophysiological and bioenergetic parameters, and nutrient content were assessed. AVP and WT plant heights were strongly influenced by inoculation with P. indica, which also promoted increases in fresh and dry matter of shoot in both genotypes. This may be related with the stimulatory effect of P. indica on ecophysiological parameters, especially photosynthetic rate, stomatal conductance, intrinsic water use efficiency and carboxylation efficiency. However, there were differences between the genotypes concerning the physiological mechanisms leading to biomass increment. In WT plants, inoculation with P. indica stimulated all H+ pumps. However, in inoculated AVP plants, H+-PPase was stimulated, but P- and V-ATPases were inhibited. Fungal inoculation enhanced nutrient uptake in both shoots and roots of WT and AVP plants, compared to uninoculated plants; but among inoculated genotypes, the nutrient uptake was lower in AVP than in WT plants. These results clearly demonstrate that the symbiosis between P. indica and AVP plants did not benefit those plants, which may be related to the inefficient colonization of this fungus on the transgenic plants, demonstrating an incompatibility of this symbiosis, which needs to be further studied.

Therapeutic properties of fungal metabolites and silver nanoparticles have been well documented. While fungal metabolites have been used for centuries as medicinal drugs, potential of biogenic silver nanoparticles has recently received attention. We have evaluated the antimicrobial potential of Aspergillus terreus crude extract, silver nanoparticles and an amalgamation of both against four pathogenic bacterial strains. Antimicrobial activity of the following was evaluated – A. terreus extract, biogenic silver nanoparticles, and a mixture containing extract and nanoparticles. Four pathogenic bacteria - Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, and Bacillus cereus were used as test organisms. Phenol, flavonoid, and alkaloid content of extract were determined to understand the chemical profile of the fungus. The extract contained significantly high amounts of phenols, flavonoids, and alkaloids. The extract and biogenic silver nanoparticle exhibited significant antibacterial activity at concentrations of 10 μg/ml and 1 μg/ml, respectively. When used in combination, the extract-nanoparticle mixture showed equally potent antibacterial activity at a much lower concentration of 2.5 μg/ml extract + 0.5 μg/ml nanoparticle. Given its high antibacterial potential, the fungal extract can be a promising source of novel drug lead compounds. The extract – silver nanoparticle mixture exhibited synergism in their antibacterial efficacy. This property can be further used to formulate new age drugs.

Staphylococcus aureus, a notorious human pathogen, is a major cause of the community as well as healthcare associated infections. It can cause a diversity of recalcitrant infections mainly due to the acquisition of resistance to multiple drugs, its diverse range of virulence factors, and the ability to produce biofilm in indwelling medical devices. Such biofilm associated chronic infections often lead to increase in morbidity and mortality posing a high socio-economic burden, especially in developing countries. Since biofilm formation and antibiotic resistance function dependent on each other, detection of biofilm expression in clinical isolates would be advantageous in treatment decision. In this premise, we attempt to investigate the biofilm formation and its association with antibiotic resistance in clinical isolates from the patients visiting tertiary health care hospitals in Nepal. Bacterial cells isolated from clinical samples identified as S. aureus were examined for in-vitro biofilm production using both phenotypic and genotypic assays. The S. aureus isolates were also examined for susceptibility patterns of clinically relevant antibiotics as well as inducible clindamycin resistance using standard microbiological techniques and D-test, respectively. Among 161 S. aureus isolates, 131 (81.4%) were methicillin resistant S. aureus (MRSA) and 30 (18.6%) were methicillin sensitive S. aureus (MSSA) strains. Although a majority of MRSA strains (69.6%) showed inducible clindamycin resistance, almost all isolates (97% and 94%) were sensitive toward chloramphenicol and tetracycline, respectively. Detection of in vitro production of biofilm revealed the association of biofilm with methicillin as well as inducible clindamycin resistance among the clinical S. aureus isolates.

In the present work, novel nanotool called ‘nano-embedded fungus’ formed by impact of synergistic association of ZnO-nanorods and fungus Piriformospora indica DSM 11827, for growth of Brassica oleracea var. botrytis (Broccoli) is reported. ZnO-nanorods were synthesized by mechanical assisted thermal decomposition process and characterized by scanning electron microscopy (SEM) for morphology, X-ray diffraction for structural studies and UV-vis absorption spectroscopy for band gap determination. Nanoembedded fungus is prepared by optimizing ZnO-nanorods concentration (500 ppm) which resulted in the increased biomass of P. indica, as confirmed by dry weight method, spore count, spread plate and microscopy techniques viz. SEM and confocal microscopy. Enhancement in B. oleracea var. botrytis is reported on treatment with nanoembedded fungus. According to the authors, this is the first holistic study focusing on the impact of ZnO-nanorods in the enhancement of fungal symbiont for enhanced biomass productivity of B. oleracea plant.

Piriformospora indica is an endophytic fungus of Sebacinaceae which colonizes the roots of many plant species and confers benefits to the hosts. We demonstrate that approximately 75% of the genes, which respond to P. indica in Arabidopsis roots, differ among seedlings grown on normal phosphate (Pi) or Pi limitation conditions, and among wild-type and the wrky6 mutant impaired in the regulation of the Pi metabolism. Mapman analyses suggest that the fungus activates different signaling, transport, metabolic and developmental programs in the roots of wild-type and wrky6 seedlings under normal and low Pi conditions. Under low Pi, P. indica promotes growth and Pi uptake of wild-type seedlings, and the stimulatory effects are identical for mutants impaired in the PHOSPHATE TRANSPORTERS1;1, -1;2 and -1;4. The data suggest that the fungus does not stimulate Pi uptake, but adapts the expression profiles to Pi limitation in Pi metabolism mutants.

Arabidopsis growth and reproduction are stimulated by the endophytic fungus Piriformospora indica. The fungus produces low amounts of auxins, but the auxin levels and the expression of auxin-regulated genes are not altered in colonized roots. Also, mutants with reduced auxin levels (ilr1-1, nit1-3, tfl2, cyp79 b2b3) respond to P. indica. However, the fungus rescues the dwarf phenotype of the auxin overproducer sur1-1 by converting free auxin into conjugates, which also results in the downregulation of the auxin-induced IAA6 and the upregulation of the P. indica–induced LRR1 gene. The fungus produces relatively high levels of cytokinins, and the cytokinin levels are higher in colonized roots compared with the uncolonized controls. trans-Zeatin cytokinin biosynthesis and the CRE1/AHK2 receptor combination are crucial for P. indica–mediated growth stimulation, while mutants lacking cis-zeatin, impaired in other cytokinin receptor combinations, or containing reduced cytokinin levels respond to the fungus. Since root colonization is not affected in the cytokinin mutants, we propose that cytokinins are required for P. indica–induced growth promotion. Finally, a comparative analysis of the phytohormone mutants allows the conclusion that the response to P. indica is independent of the architecture and size of the roots.

Piriformospora indica is an endophytic fungus that colonizes the roots of many plant species, including Arabidopsis. We exposed 18-day-old Arabidopsis seedlings, which were either cocultivated with the fungus or mock-treated for the last 9 days, to mild drought stress for 84 h. During the first 36 to 48 h, seedlings cocultivated with the fungus continued to grow, while the uncolonized controls did not. This results in a threefold difference in the fresh weight and a more than twofold difference in the chlorophyll content. The photosynthetic efficiency was only slightly reduced in the colonized (F variable/F maximum [Fv/Fm] at t0h = 0.82 and t36h = 0.79) and was severely impaired in the uncolonized (Fv/Fm at t0h = 0.81 and t36h = 0.49) seedlings, which also showed symptoms of withering. When seedlings exposed to drought stress for 72 or 84 h were transferred to soil, 10% (72 h) and none (84 h) of uncolonized seedlings reached the flowering stage and produced seeds, while 59% (72 h) and 47% (84 h) of the colonized seedlings flowered and produced seeds. After exposure to drought stress for 3 h, the message levels for RESPONSE TO DEHYDRATION 29A, EARLY RESPONSE TO DEHYDRATION1, ANAC072, DEHYDRATION-RESPONSE ELEMENT BINDING PROTEIN2A, SALT-, AND DROUGHT-INDUCED RING FINGER1, phospholipase Dδ, CALCINEURIN B-LIKE PROTEIN (CBL)1, CBL-INTERACTING PROTEIN KINASE3, and the histone acetyltransferase (HAT) were upregulated in the leaves of P. indica–colonized seedlings. Uncolonized seedlings responded 3 to 6 h later, and the message levels increased much less. We identified an Arabidopsis ethylmethane-sulfonate mutant that is less resistant to drought stress and in which the stress-related genes were not upregulated in the presence of P. indica. Thus, P. indica confers drought-stress tolerance to Arabidopsis, and this is associated with the priming of the expression of a quite diverse set of stress-related genes in the leaves. Transfer to soil was again associated with a faster and stronger upregulation of the message levels for phospholipase Dδ, CBL1, and HAT in P. indica–colonized seedlings, indicating that this response might also contribute to better survival on soil.

Endophytic actinobacteria colonize internal tissues of their host plants and are considered as a rich and reliable source of diverse species and functional microorganisms. In this study, endophytic actinobacterial strain YIM 63111 was isolated from surface-sterilized tissue of the medicinal plant Artemisia annua. We identified strain YIM 63111 as a member of the genus Pseudonocardia. A. annua seedlings grown under both sterile and greenhouse conditions were inoculated with strain YIM 63111. The growth of A. annua seedlings was strongly reduced when YIM 63111 was inoculated at higher concentrations under sterile conditions. However, no growth inhibition was observed when A. annua was grown under greenhouse conditions. Using an enhanced green fluorescent protein (EGFP) expressing YIM 63111 strain, we also observed the endophytic colonization of A. annua seedling using confocal laser-scanning microscopy. The transcription levels of the key genes involved in artemisinin biosynthesis were investigated using real time RT-PCR, revealing that cytochrome P450 monooxygenase (CYP71AV1) and cytochrome P450 oxidoreductase (CPR) expression were up-regulated in A. annua upon inoculation with strain YIM 63111 under certain conditions. The up-regulation of these genes was associated with the increased accumulation of artemisinin. These results suggest that endophytic actinobacteria effectively stimulate certain plant defense responses. Our data also demonstrate the use of Pseudonocardia sp. strain YIM 63111 as a promising means to enhance artemisinin production in plants.