Antimicrobial activity of the biologically synthesized zinc oxide nanoparticles against important rice pathogens.

Global rice production is seriously affected by many abiotic and biotic factors. Among the aggressive rice pathogens, Xanthomonas oryzae pv. oryzae (X. o. pv. oryzae), Bipolaris oryzae (B. oryzae) and Sphaerulina oryzina (S. oryzina) cause bacterial leaf blight, brown leaf spot and narrow brown leaf spot diseases, respectively. The objective of this study was to evaluate the efficacy of biogenic zinc oxide nanoparticles (ZnO NPs) as antimicrobial agent to control rice pathogens. This is the first report of antifungal activity evaluation of ZnO NPs against B. oryzae and S. oryzina. A pre-characterized bacterial strain Escherichia sp. SINT7 was bio-prospected for synthesis of green ZnO NPs. The NPs were confirmed by a characteristic peak measured at 360.96 nm through UV-Vis spectroscopy. Further, the NPs were characterized to elucidate the surface capping molecules, crystallite structure and morphology by various spectroscopic and imaging techniques, which confirmed the spherical shape of NPs with size ranging from 13.07 to 22.25 nm. In vitro studies against X. o. pv. oryzae pathogen depicted the substantial antibacterial activity (up to 25.7 mm inhibition zone at 20 µg/ml NPs concentration). Similarly, ZnO NPs reduced the mycelial growth of B. oryzae and S. oryzina up to 72.68 and 95.78%, respectively at 50 µg/ml concentration on potato dextrose agar plates, while the mycelial biomass reduction was found to be 64.66 and 68. 49% for B. oryzae and S. oryzina, respectively on potato dextrose broth media as compared to control without the addition of NPs. The green ZnO NPs also significantly reduced the fungal spore germination and a disintegration of fungal hyphae for both fungal strains was observed under the microscope as a result of NPs treatment. Hence, it was concluded that biologically synthesized ZnO NPs are potential antimicrobials and could be compared in greenhouse pathogenicity assays with commercial pesticides to control rice pathogens. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01251-y.

Bioinspired green synthesis of silver nanoparticles by using a native Bacillus sp. strain AW1-2: Characterization and antifungal activity against Colletotrichum falcatum Went.

Zero-valent silver nanoparticles (ZV-AgNPs) are known as potential antimicrobials and here we report antifungal activity of ZV-AgNPs against Colletotrichum falcatum Went for the first time. ZV-AgNPs were synthesized by using a native Bacillus sp. strain AW1-2, which was identified through 16S rRNA gene sequence analysis. Biogenic ZV-AgNPs were confirmed by monitoring a characteristic absorption peak of UV-vis spectroscopy that was measured at 447 nm. Further, it was found through FTIR and XRD analysis that ZV-Ag nanocrystals were capped with proteins of bacterial origin and their size ranged from 22.33-41.95 nm. The ultrastructure imaging through scanning electron microscopy (SEM) and transmission electron microscopy (TEM) confirmed the morphology of ZV-AgNPs as mono-dispersed spheres and energy dispersive X-ray spectroscopy (EDX) revealed the dominance of silver (84.21 %) in the nano-powder. The ZV-AgNPs significantly inhibited the hyphal growth of Colletotrichum falcatum Went as compared to non-treated control and commercial fungicide both in solid and broth media. The ultrastructure SEM and TEM studies revealed the disrupted hyphal structure and damage to the internal cellular organelles of Colletotrichum falcatum Went treated with 20 µg mL-1 ZV-AgNPs, respectively. It was concluded that green ZV-AgNPs of bacterial origin could be used to formulate a nano-based fungicide to effectively control Colletotrichum falcatum Went, the causal agent of red rot of sugarcane.

Heterologous expression of azoreductase-encoding gene azrS of Bacillus sp. MR-1/2 for enhanced azo dye decolorization and wastewater treatment.

In Pakistan, 55% of textile exports are contributed by textile-units of Faisalabad. The effluents of these textile units, being discharged without any treatment, contain the contamination of a huge amount of synthetic azo dyes. The objective of the current research was to evaluate the contribution of an azoreductase-encoding gene (azrS) from a pre-characterized azo dye decolorizing bacterial strain Bacillus sp. MR-1/2 in a high copy number host system (pUC19-T7-Top-T) of Escherichia coli strain DH5α followed by in-silico prediction of azoreductase enzyme (AzrS) function. The recombinant cells that contained azrS had a significantly higher rate of color removal in congo red and reactive black-5 dyes when compared to wild-type MR-1/2 and E. coli DH5α after 72 h of incubation. Moreover, we were able to show that the recombinant strain significantly reduced the values of all tested parameters (pH, EC, turbidity, TSS, and COD) in actual wastewater. In support of our results, it was also predicted through bioinformatics analysis that the deduced azoreductase protein of strain MR-1/2 is linked with the dye decolorization ability of the strain through NAD(P)H-ubiquinone: oxidoreductase activity. Furthermore, we also found that the deduced protein resembled closely related proteins of protein databank in many features, yet some unique features were predicted in the enzyme activity of strain MR-1/2. It was concluded that the recombinant strain could be examined in pilot-scale experiments for textile wastewater treatment.