Plant viral diseases are the foremost threat to sustainable agriculture, leading to several billion dollars in losses every year. Many viruses infecting several crops have been described in the literature; however, new infectious viruses are emerging frequently through outbreaks. For the effective treatment and prevention of viral diseases, there is great demand for new techniques that can provide accurate identification on the causative agents. With the advancements in biochemical and molecular biology techniques, several diagnostic methods with improved sensitivity and specificity for the detection of prevalent and/or unknown plant viruses are being continuously developed. Currently, serological and nucleic acid methods are the most widely used for plant viral diagnosis. Nucleic acid-based techniques that amplify target DNA/RNA have been evolved with many variants. However, there is growing interest in developing techniques that can be based in real-time and thus facilitate in-field diagnosis. Next-generation sequencing (NGS)-based innovative methods have shown great potential to detect multiple viruses simultaneously; however, such techniques are in the preliminary stages in plant viral disease diagnostics. This review discusses the recent progress in the use of NGS-based techniques for the detection, diagnosis, and identification of plant viral diseases. New portable devices and technologies that could provide real-time analyses in a relatively short period of time are prime important for in-field diagnostics. Current development and application of such tools and techniques along with their potential limitations in plant virology are likewise discussed in detail.

The stability and reusability of soluble enzymes are of major concerns, which limit their industrial applications. Herein, alkaline protease from Bacillus sp. NPST-AK15 was immobilized onto hollow core-mesoporous shell silica (HCMSS) nanospheres. Subsequently, the properties of immobilized proteases were evaluated. Non-, ethane- and amino-functionalized HCMSS nanospheres were synthesized and characterized. NPST-AK15 was immobilized onto the synthesized nano-supports by physical and covalent immobilization approaches. However, protease immobilization by covalent attachment onto the activated HCMSS–NH₂ nanospheres showed highest immobilization yield (75.6%) and loading capacity (88.1 μg protein/mg carrier) and was applied in the further studies. In comparison to free enzyme, the covalently immobilized protease exhibited a slight shift in the optimal pH from 10.5 to 11.0, respectively. The optimum temperature for catalytic activity of both free and immobilized enzyme was seen at 60 °C. However, while the free enzyme was completely inactivated when treated at 60 °C for 1 h the immobilized enzyme still retained 63.6% of its initial activity. The immobilized protease showed higher V_max, k_cat and k_cat/K_m, than soluble enzyme by 1.6-, 1.6- and 2.4-fold, respectively. In addition, the immobilized protease affinity to the substrate increased by about 1.5-fold. Furthermore, the enzyme stability in various organic solvents was significantly enhanced upon immobilization. Interestingly, the immobilized enzyme exhibited much higher stability in several commercial detergents including OMO, Tide, Ariel, Bonux and Xra by up to 5.2-fold. Finally, the immobilized protease maintained significant catalytic efficiency for twelve consecutive reaction cycles. These results suggest the effectiveness of the developed nanobiocatalyst as a candidate for detergent formulation and peptide synthesis in non-aqueous media.

Seed-borne fungus Penicillium duclauxii was examined in this study to investigate its capability of synthesizing silver nanoparticles (Ag-NPs). In vitro experiments were conducted using corn-grain contaminating fungal isolate. Ag-NPs detection and characterization were assayed by the aid of spectroscopic techniques. Spectroscopy (energy dispersive), X-ray diffraction, transmission electron-microscope and optical absorption dimensions were employed. Ag-NPs with biosynthesized were used to test invitro against Bipolaris sorghicola; the cause of target leaf spot disease on sorghum plants. The myco-synthesis of Ag NPs using Penicillium duclauxii was proved in this study. Moreover, Bipolaris sorghicola was successfully inhibited by such Ag NPs in vitro.

The present study aimed to explore the phytochemical profile, and evaluate the antioxidant, antimicrobial, and insecticidal properties, of Moroccan Mentha longifolia L. essential oil (ML-EO) using in vitro and in silico assays. Noteworthily, as chromatography (GC-MS/MS) revealed that ML-EO is majorly composed of piperitenone oxide (53.43%), caryophyllene (20.02%), and (−) germacrene D (16.53%). It possesses excellent antioxidant activity with an IC₅₀ of 1.49 ± 0.00 for DPPH and 0.051 ± 0.06 μg/mL for ABTS. Moreover, the RP and TAC activities were 0.80 ± 0.01 μg/mL and 315.532 ± 0.00 mg EAA/g, respectively. ML-EO exhibited a potent antimicrobial effect, specifically against Pseudomonas aeruginosa. It also exhibited strong antifungal ability, especially against Candida albicans. Regarding insecticidal activity, for ML-EO, a dose of 20 µL/mL produced a complete reduction in fecundity, fertility, and emergence of adult C. maculatus with mortality rates reaching 100%. In silico results showed that the antioxidant activity is mostly attributed to α-Cadinol, the antibacterial efficiency is attributed to piperitenone oxide, and antifungal capacity is related to cis-Muurola-4(15),5-diene and piperitenone oxide. Accordingly, ML-EO has high potential to be used as an alternative for preserving food and stored grain and protecting them against microbes and insect pests in the food and pharmaceutical sectors.

The drug resistance of bacterial pathogens causes considerable morbidity and death globally, hence there is a crucial necessity for the development of effective antibacterial medicines to address the antibacterial resistance issue. The bioprepared zinc oxide nanoparticles (ZnO-NPs) were prepared utilizing the flower extract of Hibiscus sabdariffa and then characterized using different physicochemical techniques. The antibacterial effectiveness of the bioprepared ZnO-NPs and their synergism with fosfomycin were evaluated using disk diffusion assay against the concerned pathogens. Transmission electron microscopy (TEM) investigation of the bioprepared ZnO-NPs showed that their average particle size was 18.93 ± 2.65 nm. Escherichia coli expressed the highest sensitivity to the bioinspired ZnO-NPs with a suppressive zone of 22.54 ± 1.26 nm at a concentration of 50 µg/disk, whereas the maximum synergistic effect of the bioinspired ZnO-NPs with fosfomycin was noticed against Klebsiella pneumoniae strain with synergism ratio of 100.29%. In conclusion, the bioinspired ZnO-NPs demonstrated significant antibacterial and synergistic efficacy with fosfomycin against the concerned nosocomial bacterial pathogens, highlighting the potential of using the ZnO NPs-fosfomycin combination for effective control of nosocomial infections in intensive care units (ICUs) and health care settings. Furthermore, the biogenic ZnO-NPs’ potential antibacterial action against food pathogens such as Salmonella typhimurium and E. coli indicates their potential usage in food packaging applications.