Deciphering the mechanistic role of Bacillus paramycoides (PM51) and Bacillus tequilensis (PM52) in bio-sorption and phyto-assimilation of Cadmium via Linum usitatissimum L. Seedlings.

Three Cd2+ resistant bacterium's minimal inhibition concentrations were assessed and their percentages of Cd2+ accumulation were determined by measurements using an atomic absorption spectrophotometer (AAS). The results revealed that two isolates Bacillus paramycoides (PM51) and Bacillus tequilensis (PM52), identified by 16S rDNA gene sequencing, showed a higher percentage of Cd2+ accumulation i.e., 83.78% and 81.79%, respectively. Moreover, both novel strains can tolerate Cd2+ levels up to 2000 mg/L isolated from district Chakwal. Amplification of the czcD, nifH, and acdS genes was also performed. Batch bio-sorption studies revealed that at pH 7.0, 1 g/L of biomass, and an initial 150 mg/L Cd2+ concentration were the ideal bio-sorption conditions for Bacillus paramycoides (PM51) and Bacillus tequilensis (PM52). The experimental data were fit to Langmuir isotherm measurements and Freundlich isotherm model R2 values of 0.999 for each of these strains. Bio sorption processes showed pseudo-second-order kinetics. The intra-diffusion model showed Xi values for Bacillus paramycoides (PM51) and Bacillus tequilensis (PM52) of 2.26 and 2.23, respectively. Different surface ligands, was investigated through Fourier-transformation infrared spectroscopy (FTIR). The scanning electron microscope SEM images revealed that after Cd2+ adsorption, the cells of both strains became thick, adherent, and deformed. Additionally, both enhanced Linum usitatissimum plant seed germination under varied concentrations of Cd2+ (0 mg/L, 250 mg/L,350 mg/L, and 500 mg/L). Current findings suggest that the selected strains can be used as a sustainable part of bioremediation techniques.

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.

Methyl eugenol aromatherapy: a delivery system facilitating the simultaneous application of male annihilation and sterile insect technique against the peach fruit fly.

BACKGROUND: The peach fruit fly, Bactrocera zonata (Saunders) (Diptera: Tephritidae) is an economically important polyphagous, quarantine pest endemic to South and South-East Asia. The male annihilation technique (MAT) and the sterile insect technique (SIT) are environmentally benign techniques used to suppress fruit fly populations on an area-wide basis. The MAT and SIT are typically used sequentially to avoid killing released sterile males; however, MAT and SIT potentially could be used simultaneously and thereby increase the overall efficiency of control programmes. Mating competitiveness of sterile males against wild counterparts is critical for the success of the SIT. Feeding on a semiochemical, methyl eugenol (ME) has been reported to enhance the male mating performance of many Bactrocera spp., including B. zonata, but its use in SIT operational programmes is limited owing to the absence of a viable delivery system. RESULTS: In the present study, we demonstrated that ME aromatherapy, a practical method for large-scale delivery of ME olfactorily, enhances the mating success of treated B. zonata males. ME aromatherapy application to 5-day-old immature males for a duration of 5 h resulted in increased mating success of males tested when sexually mature, compared to untreated males. The ME-aromatized males also exhibited reduced attraction to ME-lure. CONCLUSION: A practical delivery system for applying ME by aromatherapy to mass-reared males was developed. ME-aromatherapy enhanced male mating success and suppressed their subsequent attraction to ME, thus enabling the application of MAT and SIT at the same time. © 2023 Society of Chemical Industry.

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.

Concurrent application of bacterial-mediated and mycosynthesized ZnO nanofungicides to maintain high ascorbic acid and delay postharvest decay of apricot.

Quality of apricot fruit is affected by different biotic stresses during growth, harvesting and storage. Due to fungal attack, huge losses of its quality and quantity are observed. The present research was designed for the diagnoses and management of postharvest rot disease of apricot. Infected apricot fruit were collected, and the causative agent was identified as A. tubingensis. To control this disease, both bacterial-mediated nanoparticles (b-ZnO NPs) and mycosynthesized nanoparticles (f-ZnO NPs) were used. Herein, biomass filtrates of one selected fungus (Trichoderma harzianum) and one bacterium (Bacillus safensis) were used to reduce zinc acetate into ZnO NPs. The physiochemical and morphological characters of both types of NPs were determined. UV-vis spectroscopy displayed absorption peaks of f-ZnO NPs and b-ZnO NPs at 310-380 nm, respectively, indicating successful reduction of Zinc acetate by the metabolites of both fungus and bacteria. Fourier transform infrared (FTIR) determined the presence of organic compounds like amines, aromatics, alkenes and alkyl halides, on both types of NPs, while X-ray diffraction (XRD) confirmed nano-size of f-ZnO NPs (30 nm) and b-ZnO NPs (35 nm). Scanning electron microscopy showed flower-crystalline shape for b-ZnO NPs and spherical-crystalline shape for f-ZnO NPs. Both NPs showed variable antifungal activities at four different concentrations (0.25, 0.50, 0.75 and 1.00 mg/ml). Diseases control and postharvest changes in apricot fruit were analyzed for 15 days. Among all treatments, 0.50 mg/ml concentration of f-ZnO NPs and 0.75 mg/ml concentration of b-ZnO NPs exhibited the strongest antifungal activity. Comparatively, f-ZnO NPs performed slightly better than b-ZnO NPs. Application of both NPs reduced fruit decay and weight, maintained higher ascorbic acid contents, sustained titratable acidity, and preserved firmness of diseased fruit. Our results suggest that microbial synthesized ZnO NPs can efficiently control fruit rot, extend shelf life, and preserve the quality of apricot.

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.

Exploration of plant growth promoting traits and regulatory mechanisms of Bacillus anthracis PM21 in enhancing salt stress tolerance in maize.

Bacillus species have been reported to reduce the negative effects of salt stress on plants; the involvement of Bacillus anthracis PM21 and the internal mechanisms involved in this process are unclear. The effects of PM21 inoculation on maize plants under salt stress were investigated in this study. The study aimed to assess the ability of Bacillus anthracis PM21 to endure high levels of salinity stress while preserving the concentration of plant growth regulators. The biomass, photosynthetic pigments, relative water content (RWC), antioxidants, osmoprotectants, inorganic ion contents, regulation of plant hormones and expression of antioxidants enzyme encoded genes were investigated under normal and salinity stress conditions. Bacillus anthracis PM21 produced a significant amount of 1-aminocyclopropane-1-carboxylate deaminase enzyme (ACC deaminase) and exopolysaccharides (EPS) under salt stress and normal conditions. PM21 also produced plant growth stimulants including indole acetic acid, gibberellic acid (GA3), kinetin, and siderophore under salinity stress and normal conditions. Under salt stress, PM21 inoculation markedly increased plant growth indices, stimulate antioxidant enzyme mechanisms, osmoprotectants, and chlorophyll content. The use of qRT-PCR to analyze the transcription of targeted genes indicated greater expression of antioxidant-encoded genes and inferred their possible function in salinity stress tolerance. Our findings shed light on the functions of PM21 and its regulatory mechanisms in plant salt stress tolerance, as well as the importance of PM21 in this process. This study will provide a thorough analysis of the theoretical framework for adopting PM21 in agricultural production as an eco-friendly method to enhance crop growth and yield under salinity stress.

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.

Bio-fabrication of zinc oxide nanoparticles from Picea smithiana and their potential antimicrobial activities against Xanthomonas campestris pv. Vesicatoria and Ralstonia solanacearum causing bacterial leaf spot and bacterial wilt in tomato.

Due to an inevitable disadvantage of chemical or physical synthesis routes, biosynthesis approach to nanoparticles, especially metallic oxide is attractive nowadays. Metallic oxides nanoparticles present a new approach to the control of plant pathogens. ZnO nanoparticles (ZNPs) have very important role in phytopathology. In current study, biosynthesized ZNPs were tested against two devastating bacterial pathogens including Xanthomonas campestris pv. vesicatoria and Ralstonia solanacearum causing bacterial leaf spot and bacterial wilt in tomato. ZNPs were produced using a new extract from the plant Picea smithiana using an environmentally friendly, cost-effective and simple procedure. Zinc acetate was added to P. smithiana extract, stirred and heated to 200 °C. The white precipitation at the bottom were clear indication of synthesis of nanoparticles, which were further dried by subjecting them at 450 °C. X-ray diffraction pattern determined that the ZNPs had a crystallite size of about 26 nm, Fourier transform infrared spectroscopy indicated a peak between 450 and 550 cm-1 and the particle size estimated by dynamic light scattering was about 25 nm on average. Scanning electron microscopic analysis indicated that the particles were hexagonal in shape 31 nm in diameter. Antibacterial tests showed ZNPs synthesized by P. smithiana resulted in clear inhibition zones of 20.1 ± 1.5 and 18.9 ± 1.5 mm and 44.74 and 45.63% reduction in disease severity and 78.40 and 80.91% reduction in disease incidence in X. compestris pv. vesicatoria and R. solanacearum respectively at concentration of 100 µg/ml. Our findings reveal that the concentration of ZNPs was important for their efficient antibacterial activity. Overall, the biosynthesized ZNPs have been found to have effective antimicrobial activities against bacterial wilt and bacterial leaf spot in tomato.

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.

Biodegradation of monocrotophos by Brucella intermedia Msd2 isolated from cotton plant.

Widespread and inadequate use of Monocrotophos has led to several environmental issues. Biodegradation is an ecofriendly method used for detoxification of toxic monocrotophos. In the present study, Msd2 bacterial strain was isolated from the cotton plant growing in contaminated sites of Sahiwal, Pakistan. Msd2 is capable of utilizing the monocrotophos (MCP) organophosphate pesticide as its sole carbon source for growth. Msd2 was identified as Brucella intermedia on the basis of morphology, biochemical characterization and 16S rRNA sequencing. B. intermedia showed tolerance of MCP up to 100 ppm. The presence of opd candidate gene for pesticide degradation, gives credence to B. intermedia as an effective bacterium to degrade MCP. Screening of the B. intermedia strain Msd2 for plant growth promoting activities revealed its ability to produce ammonia, exopolysaccharides, catalase, amylase and ACC-deaminase, and phosphorus, zinc and potassium solubilization. The optimization of the growth parameters (temperatures, shaking rpm, and pH level) of the MCP-degrading isolate was carried out in minimal salt broth supplemented with MCP. The optimal pH, temperature, and rpm for Msd2 growth were observed as pH 6, 35 °C, and 120 rpm, respectively. Based on optimization results, batch degradation experiment was performed. Biodegradation of MCP by B. intermedia was monitored using HPLC and recorded 78% degradation of MCP at 100 ppm concentration within 7 days of incubation. Degradation of MCP by Msd2 followed the first order reaction kinetics. Plant growth promoting and multi-stress tolerance ability of Msd2 was confirmed by molecular analysis. It is concluded that Brucella intermedia strain Msd2 could be beneficial as potential biological agent for an effective bioremediation for polluted environments.

Integrated Application of Salicylic Acid and PGPRs to Control Fusarium Wilt of Chickpea.

BACKGROUND: Fusarium wilt and Ascochyta blight are the most important diseases of chickpea. The current study was designed to investigate the individual and combined effect of salicylic acid (SA) with Pseudomonas stutzeri and Pseudomonas putida to suppress Fusarium wilt and promote growth of chickpea varieties: Thal-2006 and Punjab-2008. METHODS: At the time of sowing, inoculum of Fusarium oxysporum was applied to the soil and the incidence of Fusarium wilt was recorded after 60 days. The seeds were inoculated with Pseudomonas stutzeri and Pseudomonas putida prior to sowing. Chickpea plants were treated with salicylic acid at seedling stage. RESULTS: The combination of P. stutzeri and SA significantly increased root length (166% and 145%), shoot height (50% and 47%) and shoot biomass (300% and 233%) in cv. Thal-2006 and cv. Punjab-2008, respectively, in infected plants. Similarly, the combined treatment of P. putida + SA, also enhanced the plant growth parameters of chickpea varieties. Maximum reduction in disease severity was observed in both P. stutzeri + SA (90% and 84%) and P. putida + SA (79% and 77%) treatments in cv. Thal-2006 and Punjab-2008, respectively. Both P. putida + SA and P. stutzeri + SA treatments resulted in increased leaf relative water and total protein content, peroxidase, superoxide dismutase, phenylalanine ammonia-lyase and polyphenol oxidase activities in both resistant (cv. Thal-2006) and susceptible (cv. Punjab-2008) cultivars. Both treatments also significantly reduced malondialdehyde (MDA) and proline content in cv. Thal-2006 and Punjab-2008. Cultivar Thal-2006 was more effective than cv. Punjab-2008. CONCLUSIONS: The results suggested that, in combination, salicylic acid and P. stutzeri may play an important role in controlling Fusarium wilt diseases by inducing systemic resistance in chickpea.

Hydropriming and Osmotic Priming Induce Resistance against Aspergillus niger in Wheat (Triticum aestivum L.) by Activating ß-1, 3-glucanase, Chitinase, and Thaumatin-like Protein Genes.

Priming is used as a method to improve plant growth and alleviate the detrimental effects of pathogens. The present study was conducted to evaluate the effects of different priming methods in the context of resistance to Aspergillus niger in wheat (Triticum aestivum L.). Here, we show that different priming treatments­viz., hydropriming, osmotic priming, halopriming, and hormonal priming techniques can induce disease resistance by improving the biochemical contents of wheat, including chlorophyll, protein, proline, and sugar. In addition, physiological parameters­such as root length, shoot length, fresh and dry root/shoot ratios, and relative water content were positively affected by these priming methods. In essence, hydropriming and osmotic priming treatments were found to be more potent for enhancing wheat biochemical contents, along with all the physiological parameters, and for reducing disease severity. Hydropriming and osmotic priming significantly decreased disease severity, by 70.59−75.00% and 64.71−88.33%, respectively. RT-PCR and quantitative real-time PCR analyses of potentially important pathogenesis-related (PR)-protein genes (Thaumatin-like protein (TLP), chitinase, and ß-1,3-glucanase) in primed plants were evaluated: ß-1,3-glucanase was most highly expressed in all primed plants; Chitinase and TLP exhibited higher expression in hormonal-, halo-, osmotic-, and hydro-primed plants, respectively. These results suggest that the higher expression of ß-1,3-glucanase, TLP, and chitinase after hydropriming and osmotic priming may increase disease resistance in wheat. Our study demonstrates the greater potential of hydropriming and osmotic priming for alleviating stress caused by A. niger inoculation, and enhancing resistance to it, in addition to significantly improving plant growth. Thus, these priming methods could be beneficial for better plant growth and disease resistance in other plants.

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.

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.