Identification of therapeutic drug target of Shigella Flexneri serotype X through subtractive genomic approach and in-silico screening based on drug repurposing.

Shigellosis, induced by Shigella flexneri, constitutes a significant health burden in developing nations, particularly impacting socioeconomically disadvantaged communities. Designated as the second most prevalent cause of diarrheal illness by the World Health Organization (WHO), it precipitates an estimated 212,000 fatalities annually. Within the spectrum of S. flexneri strains, serotype X is notably pervasive and resilient, yet its comprehensive characterization remains deficient. The present investigation endeavors to discern potential pharmacological targets and repurpose existing drug compounds against S. flexneri serotype X. Employing the framework of subtractive genomics, the study interrogates the reference genome of S. flexneri Serotype X (strain 2,002,017; UP000001884) to delineate its proteome into categories of non-homologous, non-paralogous, essential, virulent, and resistant constituents, thereby facilitating the identification of therapeutic targets. Subsequently, a screening of approximately 9000 compounds from the FDA library against the identified drug target aims to delineate efficacious agents for combating S. flexneri serotype X infections. The application of subtractive genomics methodology yields prognostic insights, unveiling non-paralogous proteins (n = 4122), non-homologues (n = 1803), essential (n = 1246), drug-like (n = 389), resistant (n = 167), alongside 42 virulent proteins within the reference proteome. This iterative process culminates in the identification of Serine O-acetyltransferase as a viable drug target. Subsequent virtual screening endeavors to unearth FDA-approved medicinal compounds capable of inhibiting Serine O-acetyltransferase. Noteworthy candidates such as DB12983, DB15085, DB16098, DB16185, and DB16262 emerge, exhibiting potential for mitigating S. flexneri Serotype X. Despite the auspicious findings, diligent scrutiny is imperative to ascertain the efficacy and safety profile of the proposed drug candidates vis-à-vis S. flexneri.

Bacillus safensis filtrate-based ZnO nanoparticles control black heart rot disease of apricot fruits by maintaining its soluble sugars and carotenoids.

Black heart rot is a serious disease of apricot and it has been reported to be caused by Alternaria solani, around the world. The present research was designed to control this disastrous disease using zinc oxide nanoparticles (b-ZnO NPs). These NPs were synthesized in the filtrate of a useful bacterium (Bacillus safensis) and applied to control black heart rot of apricot. After synthesis, the reduction of b-ZnO NPs was confirmed by UV-visible spectroscopy, at 330 nm. Fourier transform infrared (FTIR) spectra ensured the presence of multiple functional groups (alcohols, phenols, carboxylic acids, nitro compounds and amines) on the surface of b-ZnO NPs. X-Ray diffraction (XRD) analysis elucidated their average size (18 nm) while scanning electron microscopy (SEM) micrograph described the spherical shape of b-ZnO NPs. The synthesized b-ZnO NPs were applied in four different concentrations (0.25 mg/ml, 0.50 mg/ml, 0.75 mg/ml, 1.0 mg/ml) under both in vitro and in vivo conditions. These NPs were very efficient in inhibiting mycelial growth (85.1%) of A. solani at 0.75 mg/ml concentration of NPs, in vitro. Same concentration also performed best, in vivo, and significantly reduced disease incidence (by 67%) on self-inoculated apricot fruit. Apart from this, application of b-ZnO NPs helped apricot fruit to maintain its quality under fungal-stress conditions. The decay of apricot fruit was reduced and they maintained greater firmness and higher weight. Moreover, b-ZnO NPs treated fruits controlled black heart rot disease by maintaining higher contents of ascorbic acid, soluble sugars and carotenoids. These b-ZnO NPs were produced in powder form for their easy carriage to the farmers' fields.

Spherical Fe2O3 nanoparticles inhibit the production of aflatoxins (B1 and B2) and regulate total soluble solids and titratable acidity of peach fruit.

Aflatoxin is a group I carcinogen and causes significant public health and food safety risks, throughout the world. This study was carried out to assess the levels of aflatoxin contamination in diseased peach (Prunus persica L.) fruit and their control using myco-synthesized iron oxide nanoparticles (Fe2O3 NPs). Diseased peach fruit were diagnosed to be infected with Aspergillus flavus. The isolated pathogen was cultured under UV light (365 nm) and exposed to ammonium hydroxide (31 %) vapors, which confirmed its ability to produce aflatoxin. For the control of this disease, Fe2O3 NPs were synthesized in the filtrate of a biocontrol fungus (Trichoderma harzianum) and characterized before analyzing their potential in disease control. FTIR spectrum described the presence of capping and reducing agents (secondary amines, alcohol, alkyne and aromatic compounds) on the surface of Fe2O3 NPs. X-ray Diffraction (XRD) described the crystalline size (7.78), while the spherical shape of Fe2O3 NPs was described by the SEM analysis. The EDX spectrum indicated the successful formation of Fe2O3 NPs by showing strong signals of iron (74.38 %). All concentrations displayed mycelial growth inhibition, in vitro and the greatest growth reduction (65.4 %) was observed at 1 mg/ml concentration of NPs. At the same concentration of Fe2O3 NPs, significant control of fruit rot of peach was also observed, in vivo. Treatment of Fe2O3 NPs maintained higher soluble solids, sucrose, total sugar, ascorbic acid, titratable acidity and firmness of peach fruit. Diseased fruit were further investigated for the presence and detection of aflatoxins. All three methods viz. thin layer chromatography (TLC), enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC) confirmed a higher production of aflatoxins in control plants, while this production was significantly reduced in Fe2O3 NPs-treated peach fruit.

Impact of bacterial synthesized nanoparticles on quality attributes and postharvest disease control efficacy of apricot and loquat.

Postharvest fungal attacks on fruits such as apricots and loquats are common. Diseased fruit samples were collected from Murree's local fruit markets. The disease-causing pathogens were identified utilizing molecular, microscopic, and morphological characteristics. Alternaria alternata and Aspergillus niger were identified as the pathogens responsible for brown rot in loquat and black rot in apricot. To combat these fruit diseases, iron oxide (Fe2 O3 ) nanoparticles were synthesized using Bacillus subtilis and were characterized using various techniques. X-ray diffraction examination validated the size of iron oxide nanoparticles. The presence of several capping agents in the synthesized nanoparticles was confirmed by Fourier transform infrared analysis. Scanning electron microscopy revealed the spherical morphology of nanoparticles, whereas energy-dispersive X-ray proved the presence of different elemental compositions. After completing antifungal activities in vitro and in vivo, it was discovered that a nanoparticle concentration of 1.0 mg/mL efficiently suppressed the growth of fungal mycelia. Fungi growth was effectively inhibited in fruit samples treated with 1.0 mg/mL nanoparticles. The results of successful in vitro and in vivo antifungal activities imply that iron oxide (Fe2 O3 ) nanoparticles play an important role in ensuring fruit quality against pathogenic attacks. Bacterial-mediated iron oxide can be widely used because it is less expensive and less harmful to the environment than chemically manufactured fertilizers.

Effect of green Fe2O3 nanoparticles in controlling Fusarium fruit rot disease of loquat in Pakistan.

The subtropical fruit known as the loquat is prized for both its flavour and its health benefits. The perishable nature of loquat makes it vulnerable to several biotic and abiotic stressors. During the previous growing season (March-April 2021), loquat in Islamabad showed signs of fruit rot. Loquat fruits bearing fruit rot symptoms were collected, and the pathogen that was causing the disease isolated and identified using its morphology, microscopic visualisation, and rRNA sequence. The pathogen that was isolated was identified as Fusarium oxysporum. Green synthesized metallic iron oxide nanoparticles (Fe2O3 NPs) were employed to treat fruit rot disease. Iron oxide nanoparticles were synthesized using a leaf extract of the Calotropis procera. Characterization of NPs was performed by different modern techniques. Fourier transform infrared spectroscopy (FTIR) determined the existence of stabilizing and reducing compounds like phenol, carbonyl compounds, and nitro compounds, on the surface of Fe2O3 NPs. X-ray diffraction (XRD) explained the crystalline nature and average size (~49 nm) of Fe2O3 NPs. Energy dispersive X-ray (EDX) exhibited Fe and O peaks, and scanning electron microscopy (SEM) confirmed the smaller size and spherical shape of Fe2O3 NPs. Following both in vitro and in vivo approaches, the antifungal potential of Fe2O3 NPs was determined, at different concentrations. The results of both in vitro and in vivo analyses depicted that the maximum fungal growth inhibition was observed at concentration of 1.0 mg/mL of Fe2O3 NPs. Successful mycelial growth inhibition and significantly reduced disease incidence suggest the future application of Fe2O3 NPs as bio fungicides to control fruit rot disease of loquat.

Application of phyto-fabricated zinc oxide nanoparticles to diminish brown spot of pear whilst maintaining its quality.

Pyrus communis is a common fruit of temperate region, its domestication and origin is at two different points, Asia and China. Pear fruits bearing brown spot symptoms were picked, and pathogen was isolated and poured on Potato Dextrose Agar (PDA) media. On basis of molecular and molecular analyses, this isolated pathogen was identified as Alternaria alternata. Zinc oxide nanoparticles (ZnO NPs) were prepared from Theveti peruviana leaf extract and were characterized through various techniques Fourier Transform Infrared Spectroscopy (FTIR) analysis of prepared ZnO NPs depicted the presence of agents responsible for stabilization and reduction such as alkenes, alkynes, nitro compounds, alkyl halides, aromatic compounds and aliphatic amines. X-ray diffraction (XRD) analysis confirmed the size (27 nm) and crystalline nature of ZnO NPs. Scanning electron microscopy (SEM) depicted the irregular shape of the prepared ZnO NPs. Mass percentage of zinc (79.84%) and oxygen (20.16%) was depicted using Energy Dispersive X-Ray (EDX) analysis. The in vitro and in vivo antifungal activity (A.F) of prepared NPs against A. alternata was confirmed by poisoned food technique and wound inoculation method. On the basis of which it was concluded that 1.0 mg/mL concentration of ZnO NPs could effectively inhibit A. alteranata growth and minimize the risk of brown spot of pear. SEM images of A. alternata under 1 mg/mL NPs showed the deformation in morphology of A. alternata. ZnO NPs also aided in the preservation of its various organoleptic and biochemical properties. The high percentage of soluble solids, firmness, ascorbic acid and sugars demonstrated its high quality. It has been concluded that 1 mg/mL ZnO NPs can effectively control brown spot of pear while maintaining its quality. In addition, the method might be applied to control emerging diseases in an ecofriendly way to meet the global food demand. RESEARCH HIGHLIGHTS: Isolation and characterization of pathogen causing brown spot in pear. Pathogenicity of A. alternata was checked on healthy fruits. Thevetia peruviana leaf extract was used for the synthesis, characterization and antifungal assay of ZnO Nanoparticles. Green synthesized nanoparticles can be economically effective alternative fungicide for the large scale in agriculture fields.

Biofilm formation and flocculation potential analysis of halotolerant Bacillus tequilensis and its inoculation in soil to mitigate salinity stress of chickpea.

Application of beneficial microbes in soil is an important avenue to control plant stresses. In this study, the salinity tolerance of halotolerant bacteria (Bacillus tequilensis) was investigated and the bacterium was inoculated in the soil to mitigate salinity stress. The results revealed the highest floc yield and biofilm formation ability of B. tequilensis at 100 mM NaCl concentration. Fourier transformed infrared spectroscopy depicted the presence of carbohydrates and proteins which binds with sodium ions (Na+) and provide tolerance against salinity. Using PCR, plant growth-promoting bacterial genes viz., 1-aminocyclopropane-1-carboxylate deaminase and pyrroloquinoline quinone were successfully amplified from the genome of B. tequilensis. In the saline soil, B. tequilensis was inoculated and chickpea plants were grown. The bacterial strain improved the physiology, biochemistry, and antioxidant enzyme activities of the chickpea plant under salt stress. Plants inoculated with B. tequilensis exhibited higher relative water content, higher photosynthetic pigments, lower levels of hydrogen peroxide (H2O2) and malondialdehyde, and improved enzymatic activity for the scavenging of reactive oxygen species. The findings of this study suggest the sustainable use of B. tequilensis to mitigate the salinity stress of chickpea and other crops. This bacterium not only helps in the alleviation of the toxic effects of salt but also increases plant growth along with a reduction in crop losses due to salinity. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01280-1.

Development, validation and forced degradation studies of green liquid chromatographic method for determination of remdesivir in bulk drug and pharmaceutical formulation.

Remdesivir was recently approved by Food and Drug Administration (FDA) to treat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease. In this study, we report development of the green and reversed-phase liquid chromatographic method for quantitative determination of remdesivir in pharmaceutical formulations in forced degradation studies using mobile phase consisting of 0.4% trifluoroacetic acid: Acetonitrile with flow rate of 1.0mL min-1. High performance liquid chromatographic stability-indicating procedure was evaluated and impurities were profiled by subjecting the remdesivir under stress conditions i.e. acidic and alkaline hydrolysis, oxidative and thermal degradation. The developed method effectively separated the parent drug response from that of degradation products and it was validated following ICH guidelines within linearity range of 5-50µg mL-1 exhibiting correlation coefficient greater than 0.997. At all levels, the % RSD values were less than 2.0 indicating satisfactory precision. Green profile of developed method was evaluated by NEMI and AGREE tools. Comparison of proposed method with already existing chromatographic method was established using analytical eco-scale. The proposed method is eco-friendly and resulted in reliable quantification of remdesivir in dosage formulation and impurity profiling.

Reprisal of Schima superba to Mn stress and exploration of its defense mechanism through transcriptomic analysis.

One of the most diverse protein families, ATP-binding cassette (ABC) transporters, play a role in disease resistance, heavy metal tolerance, and food absorption.Differentially expressed genes contribute in the investigation of plant defense mechanisms under varying stress conditions. To elucidate the molecular mechanisms involved in Mn metal stress, we performed a transcriptomic analysis to explore the differential gene expression in Schima superba with the comparison of control. A total of 79.84 G clean data was generated and 6558 DEGs were identified in response to Mn metal stress. Differentially expressed genes were found to be involved in defense, signaling pathways, oxidative burst, transcription factors and stress responses. Genes important in metal transport were more expressive in Mn stress than control plants. The investigation of cis-acting regions in the ABC family indicated that these genes might be targeted by a large variety of trans-acting elements to control a variety of stress circumstances. Moreover, genes involved in defense responses, the mitogen-activated protein kinase (MAPK) signaling and signal transduction in S. superba were highly induced in Mn stress. Twenty ABC transporters were variably expressed on 1st, 5th, and 10th day of Mn treatment, according to the qRT PCR data. Inclusively, our findings provide an indispensable foundation for an advanced understanding of the metal resistance mechanisms. Our study will enrich the sequence information of S. superba in a public database and would provide a new understanding of the molecular mechanisms of heavy metal tolerance and detoxification.

Biocontrol of Fruit Rot of Litchi chinensis Using Zinc Oxide Nanoparticles Synthesized in Azadirachta indica.

Lychee (Litchi chinensis Sonn.) is a famous fruit species of tropical and subtropical regions of the world and many biotic and abiotic stresses affect its yield. In this study, lychee fruit rot has been observed and its incidence has been controlled by using zinc oxide nanoparticles (ZnO NPs). Diseased lychee fruits were collected and diagnosed to identify disease-causing pathogens. Morphological appearance, microscopic observation, and sequence analysis of the amplified ITS region identified this isolated pathogen as Aspergillus niger. To control this problem, ZnO NPs were prepared in the leaf extract of Azadirachta indica. Before their antifungal activity, ZnO NPs were characterized using sophisticated approaches. FTIR revealed the presence of reducing and stabilizing molecules on ZnO NPs including alcohol, carboxylic acid, alkyl halide, amine, and alkyl halide. Crystalline nature and average size (29.024 nm) of synthesized ZnO NPs were described by X-ray diffraction. EDX analysis depicted the mass percentage of zinc (30.15%) and oxygen (14.90%). SEM analysis displayed the irregular shape of nanoparticles and confirmed the nano-size of ZnO NPs. Maximum mycelial growth inhibition (70.5%) was observed at 1.0 mg/mL concentration of ZnO NPs in vitro. In in-vivo disease-control analysis, maximum control of lychee fruit rot disease was observed at the same concentration. These results reveal the potential use of these ZnO NPs on a larger scale to replace hazardous chemical fungicides.

Biological Applications of Ball-Milled Synthesized Biochar-Zinc Oxide Nanocomposite Using Zea mays L.

Nanotechnology is one of the vital and quickly developing areas and has several uses in various commercial zones. Among the various types of metal oxide-based nanoparticles, zinc oxide nanoparticles (ZnO NPs) are frequently used because of their effective properties. The ZnO nanocomposites are risk-free and biodegradable biopolymers, and they are widely being applied in the biomedical and therapeutics fields. In the current study, the biochar-zinc oxide (MB-ZnO) nanocomposites were prepared using a solvent-free ball-milling technique. The prepared MB-ZnO nanocomposites were characterized through scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) spectroscopy, X-ray powder diffraction (XRD), and thermogravimetric analysis (TGA), Fourier-transform infrared spectroscopy (FTIR), and ultraviolet-visible (UV) spectroscopy. The MB-ZnO particles were measured as 43 nm via the X-ray line broadening technique by applying the Scherrer equation at the highest peak of 36.36°. The FTIR spectroscope results confirmed MB-ZnO's formation. The band gap energy gap values of the MB-ZnO nanocomposites were calculated as 2.77 eV by using UV-Vis spectra. The MB-ZnO nanocomposites were tested in various in vitro biological assays, including biocompatibility assays against the macrophages and RBCs and the enzymes' inhibition potential assay against the protein kinase, alpha-amylase, cytotoxicity assays of the leishmanial parasites, anti-inflammatory activity, antifungal activity, and antioxidant activities. The maximum TAC (30.09%), TRP (36.29%), and DPPH radicals' scavenging potential (49.19%) were determined at the maximum dose of 200 µg/mL. Similarly, the maximum activity at the highest dose for the anti-inflammatory (76%), at 1000 µg/mL, alpha-amylase inhibition potential (45%), at 1000 µg/mL, antileishmanial activity (68%), at 100 µg/mL, and antifungal activity (73 ± 2.1%), at 19 mg/mL, was perceived, respectively. It did not cause any potential harm during the biocompatibility and cytotoxic assay and performed better during the anti-inflammatory and antioxidant assay. MB-ZnO caused moderate enzyme inhibition and was more effective against pathogenic fungus. The results of the current study indicated that MB-ZnO nanocomposites could be applied as effective catalysts in various processes. Moreover, this research provides valuable and the latest information to the readers and researchers working on biopolymers and nanocomposites.

ZnO Nanoparticle-Mediated Seed Priming Induces Biochemical and Antioxidant Changes in Chickpea to Alleviate Fusarium Wilt.

Chickpea (Cicer arietinum L.) is one of the main pulse crops of Pakistan. The yield of chickpea is affected by a variety of biotic and abiotic factors. Due to their environmentally friendly nature, different nanoparticles are being synthesized and applied to economically important crops. In the present study, Trichoderma harzianum has been used as a stabilizing and reducing agent for the mycosynthesis of zinc oxide nanoparticles (ZnO NPs). Before their application to control Fusarium wilt of chickpea, synthesized ZnO NPs were characterized. X-ray diffraction (XRD) analysis revealed the average size (13 nm) of ZnO NPs. Scanning electron microscopy (SEM) indicated their spherical structure, and energy dispersive X-ray analysis (EDX) confirmed the oxide formation of ZnO NPs. Transmission electron microscopy (TEM) described the size and shape of nanoparticles, and Fourier transform infrared (FTIR) spectroscopy displayed the presence of reducing and stabilizing chemical compounds (alcohol, carboxylic acid, amines, and alkyl halide). Successfully characterized ZnO NPs exhibited significant mycelial growth inhibition of Fusarium oxysporum, in vitro. In a greenhouse pot experiment, the priming of chickpea seeds with ZnO NPs significantly increased the antioxidant activity of germinated plants and they displayed 90% less disease incidence than the control. Seed priming with ZnO NPs helped plants to accumulate higher quantities of sugars, phenol, total proteins, and superoxide dismutase (SOD) to create resistance against wilt pathogen. These nanofungicides were produced in powder form and they can easily be transferred and used in the field to control Fusarium wilt of chickpea.

First Report of Fruit Rot of Cherry and Its Control Using Fe2O3 Nanoparticles Synthesized in Calotropis procera.

Cherry is a fleshy drupe, and it is grown in temperate regions of the world. It is perishable, and several biotic and abiotic factors affect its yield. During April-May 2021, a severe fruit rot of cherry was observed in Swat and adjacent areas. Diseased fruit samples were collected, and the disease-causing pathogen was isolated on PDA. Subsequent morphological, microscopic, and molecular analyses identified the isolated pathogen as Aspergillus flavus. For the control of the fruit rot disease of cherry, iron oxide nanoparticles (Fe2O3 NPs) were synthesized in the leaf extract of Calotropis procera and characterized. Fourier transform infrared (FTIR) spectroscopy of synthesized Fe2O3 NPs showed the presence of capping and stabilizing agents such as alcohols, aldehydes, and halo compounds. X-ray diffraction (XRD) analysis verified the form and size (32 nm) of Fe2O3 NPs. Scanning electron microscopy (SEM) revealed the spinal-shaped morphology of synthesized Fe2O3 NPs while X-ray diffraction (EDX) analysis displayed the occurrence of main elements in the samples. After successful preparation and characterization of NPs, their antifungal activity against A. flavus was determined by poison technique. Based on in vitro and in vivo antifungal activity analyses, it was observed that 1.0 mg/mL concentration of Fe2O3 can effectively inhibit the growth of fungal mycelia and decrease the incidence of fruit rot of cherry. The results confirmed ecofriendly fungicidal role of Fe2O3 and suggested that their large-scale application in the field to replace toxic chemical fungicides.

Development and evaluation of herbal formulation AKIGTU01 and AKIGCL03 against acne producing microbes.

Acne vulgaris is a common global skin disease affecting teenagers and adults and exerting serious psychological impacts which includes everlasting scarring, reduced self-image, depression and anxiety. One of the suspected causative agent of acne is Propionibacterium acnes; a Gram positive anaerobic organism which lives in skin hair follicle and openings. Treatments currently available for acne include use of oral antibiotics, hormones, isotretinoin and also physical treatments like lesion removal and photo-therapy. All these are associated with risks and none is completely satisfactory.Therefore, natural alternatives are gaining greater research support but lacks sufficient studies. In our study we have isolated Propionibacterium acnes from infected individuals and tested the effect of certain chemicals and herbs/ vegetable extracts against it. There anti-acne property was studied and compared with commercially used antibiotics including Clinagel (Clindamycin phosphate), Vibramycin (Doxycycline), Erythromycin, Novidat (Ciprofloxacin) and Amoxil (Amoxicillin). Results indicate that some of the selected herbs and chemicals showed good activity against Propionibacterium acnes synergistic to the antibiotics when used alone or in combination. Findings of this research can play an important role in natural product based drug discovery for the treatment of Acne vulgaris.

Antifungal activity of Zinc nitrate derived nano Zno fungicide synthesized from Trachyspermum ammi to control fruit rot disease of grapefruit.

Grapefruit (Citrus paradisi) is a widely grown citrus and its fruit is affected by a variety of biotic and abiotic stress. Keeping in view the hazardous effects of synthetic fungicides, the recent trend is shifting towards safer and eco-friendly control of fruit diseases. The present study was aimed to diagnose the fruit rot disease of grapefruit and its control by using zinc oxide green nanoparticles (ZnO NPs). Fruit rot symptoms were observed in various grapefruit growing sites of Pakistan. Diseased samples were collected, and the disease-causing pathogen was isolated. Following Koch's postulates, the isolated pathogen was identified as Rhizoctonia solani. For eco-friendly control of this disease, ZnO NPs were prepared in the seed extract of Trachyspermum ammi and characterized. Fourier transform infrared spectroscopy (FTIR) of these NPs described the presence of stabilizing and reducing compounds such as phenols, aldehyde and vinyl ether, especially thymol (phenol). X-ray diffraction (XRD) analysis revealed their crystalline nature and size (48.52 nm). Energy dispersive X-ray (EDX) analysis elaborated the presence of major elements in the samples, while scanning electron microscopy (SEM) confirmed the morphology of bio fabricated NPs. ZnO NPs exhibited very good anti-fungal activity and the most significant fungal growth inhibition was observed at 1.0 mg/ml concentration of green NPs, in vitro and in vivo. These findings described that the bioactive constituents of T. ammi seed extract can effectively reduce and stabilize ZnO NPs. It is a cost-effective method to successfully control the fruit rot disease of grapefruit.

Mycosynthesized Fe2 O3 nanoparticles diminish brown rot of apple whilst maintaining composition and pertinent organoleptic properties.

AIMS: Iron oxide nanoparticles (Fe2 O3 NPs) were mycosynthesized using Trichoderma harzianum and applied to control brown rot of apple. The influence of Fe2 O3 NPs on the quality of fruit was also studied. METHODS AND RESULTS: Diseased apple fruits with brown rot symptoms were collected, and the disease-causing pathogen was isolated and identified as Fusarium oxysporum. To control this disease, mycosynthesis of Fe2 O3 NPs was executed using T. harzianum. FTIR spectroscopy revealed the occurrence of stabilizing and reducing agents on NPs. X-ray diffraction (XRD) analysis determined their average size (17.78 nm) and crystalline nature. Energy-dispersive X-ray (EDX) showed strong signals of iron, and scanning electron microscopy (SEM) displayed a high degree of polydispersity of synthesized NPs. Foliar application of NPs significantly reduced brown rot and helped fruits to maintain biochemical and organoleptic properties. Firmness and higher percentage of soluble solids, sugars and ascorbic acid depicted its good quality. CONCLUSION: Environment-friendly mycosynthesized Fe2 O3 NPs can be effectively used to control brown rot of apple. SIGNIFICANCE AND IMPACT OF THE STUDY: Trichoderma harzianum is a famous biocontrol agent, and the synthesis of NPs in its extract is an exciting avenue to control fungal diseases. Due to its nontoxic nature to human gut, it can be applied on all edible fruits.

Evaluation of virulence potential of Aspergillus tubingensis and subsequent biochemical and enzymatic defense response of cotton.

Aspergillus tubingensis is a causative known pathogen of various important crops, worldwide. The existing study was aimed to examine the virulence potential of A. tubingensis on resistant (NIA-Sadori) and susceptible (CIM-573) cultivars of cotton. For this purpose, both cultivars were inoculated with pathogen and altered morphology of diseased leaves was observed with light and scanning electron microscope. Disease severity was measured and estimated to be 68.7 and 27.1% in susceptible and resistant cultivars, respectively. To understand and compare defense mechanism of resistant and susceptible cultivars, different biochemical and enzymatic changes were observed. After the infection of A. tubingensis, increase in the concentrations of sugar, total protein, proline, phenol, and phenylalanine ammonia lyase (PAL) was more prominent in resistant cultivar, than the susceptible one. Moreover, due to increased number of dead cells, significantly higher electrolyte leakage was detected in susceptible cultivar. Principal component analysis confirmed the effect of A. tubingensis on growth attributes and various physiological and biochemical activities of cotton. These findings help us to suggest a possible role of proline content, protein content, and PAL activity in resistance mechanism of Cotton plant.

PacBio Single-Molecule Long-Read Sequencing Reveals Genes Tolerating Manganese Stress in Schima superba Saplings.

Schima superba (Theaceae) is a subtropical evergreen tree and is used widely for forest firebreaks and gardening. It is a plant that tolerates salt and typically accumulates elevated amounts of manganese in the leaves. With large ecological amplitude, this tree species grows quickly. Due to its substantial biomass, it has a great potential for soil remediation. To evaluate the thorough framework of the mRNA, we employed PacBio sequencing technology for the first time to generate S. Superba transcriptome. In this analysis, overall, 511,759 full length non-chimeric reads were acquired, and 163,834 high-quality full-length reads were obtained. Overall, 93,362 open reading frames were obtained, of which 78,255 were complete. In gene annotation analyses, the Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Genes (COG), Gene Ontology (GO), and Non-Redundant (Nr) databases were allocated 91,082, 71,839, 38,914, and 38,376 transcripts, respectively. To identify long non-coding RNAs (lncRNAs), we utilized four computational methods associated with protein families (Pfam), Cooperative Data Classification (CPC), Coding Assessing Potential Tool (CPAT), and Coding Non-Coding Index (CNCI) databases and observed 8,551, 9,174, 20,720, and 18,669 lncRNAs, respectively. Moreover, nine genes were randomly selected for the expression analysis, which showed the highest expression of Gene 6 (Na_Ca_ex gene), and CAX (CAX-interacting protein 4) was higher in manganese (Mn)-treated group. This work provided significant number of full-length transcripts and refined the annotation of the reference genome, which will ease advanced genetic analyses of S. superba.

Scanning electron microscopy of bio-fabricated Fe2 O3 nanoparticles and their application to control brown rot of citrus.

Citrus is the leading fruit crop of Pakistan and exported to different parts of the world. Due to suitable weather condition, this crop is affected by different biotic factors which seriously deteriorate its quality and quantity. During the months of November 2018 to January 2019, citrus brown rot symptoms were recurrently observed on sweet oranges in National Agricultural Research Centre (NARC), Islamabad. Causal agent of citrus brown rot was isolated, characterized, and identified as Fusarium oxysporum. For environment-friendly control of this disease, leaf extract of Azadirachta indica was used for the green synthesis of iron oxide (Fe2 O3 ) nanoparticles. These nanoparticles were characterized before their application for disease control. Fourier transform infrared spectroscopy (FTIR) of these synthesized nanoparticles described the presence of stabilizing and reducing compounds like alcohol, phenol, carboxylic acid, and alkaline and aromatic compounds. X-Ray diffraction (XRD) analysis revealed the crystalline nature and size (24 nm) of these nanoparticles. Energy dispersive X-Ray (EDX) analysis elaborated the presence of major elements in the samples. Scanning electron microscopy (SEM) confirmed the spinal shaped morphology of prepared nanoparticles. Successfully synthesized nanoparticles were evaluated for their antifungal potential. Different concentrations of Fe2 O3 nanoparticles were used and maximum mycelial inhibition was observed at 1.0 mg/ml concentration. On the basis of these findings, it could be concluded that Fe2 O3 nanoparticles, synthesized in the leaf extract of A. indica, can be successfully used for the control of brown rot of sweet oranges.

Evaluation of Metal Tolerance of Fungal Strains Isolated from Contaminated Mining Soil of Nanjing, China.

Rapidly increasing industry has resulted in greater discharge of hazardous chemicals in the soil. In the current study, soil samples were collected from Nanjing mine (32°09'19.29″ N 118°56'57.04″ E) and subjected to heavy metal analysis and microbe isolation. A total of 460 fungi were isolated, and five of these were yeast strains. Most of the strains exhibited tolerance to one metal. Five multimetal tolerant strains were selected and identified as Aspergillus sclerotiorum, Aspergillus aculeatus, Komagataella phaffii, Trichoderma harzianum, and Aspergillus niger. Isolated strains were grown in high concentrations of cadmium (Cd), chromium (Cr) and lead (Pb), for induced-tolerance training. The tolerance index (TI) revealed the highest Cd tolerance of novel K. phaffii strain at 5500 ppm (TI: 0.2). K. phaffii also displayed resistance at 4000 ppm against Cr (TI: 0.32) and Pb (TI: 0.32). In contrast, tolerance training for A. niger was not that successful. K. phaffii also displayed the highest bioaccumulation capacity for Cd (25.23 mg/g), Cu (21.63 mg/g), and Pb (20.63 mg/g) at 200 ppm. Scanning electron microscopy (SEM) explored the morphological changes in the mycelia of stressed fungi. Results of this study describe this delicate approach to be species and metal dependent and suggest a potential utilization of this fungal strain for the bioremediation of contaminated soils.