Evaluation of Phoma sp. Biomass as an Endophytic Fungus for Synthesis of Extracellular Gold Nanoparticles with Antibacterial and Antifungal Properties.

The aim of our study was to examine the different concentrations of AuNPs as a new antimicrobial substance to control the pathogenic activity. The extracellular synthesis of AuNPs performed by using Phoma sp. as an endophytic fungus. Endophytic fungus was isolated from vascular tissue of peach trees (Prunus persica) from Baft, located in Kerman province, Iran. The UltraViolet-Visible Spectroscopy (UV-Vis spectroscopy) and Fourier transform infrared spectroscopy provided the absorbance peak at 526 nm, while the X-ray diffraction and transmission electron microscopy images released the formation of spherical AuNPs with sizes in the range of 10-100 nm. The findings of inhibition zone test of Au nanoparticles (AuNPs) showed a desirable antifungal and antibacterial activity against phytopathogens including Rhizoctonia solani AG1-IA (AG1-IA has been identified as the dominant anastomosis group) and Xanthomonas oryzae pv. oryzae. The highest inhibition level against sclerotia formation was 93% for AuNPs at a concentration of 80 µg/mL. Application of endophytic fungus biomass for synthesis of AuNPs is relatively inexpensive, single step and environmentally friendly. In vitro study of the antifungal activity of AuNPs at concentrations of 10, 20, 40 and 80 µg/mL was conducted against rice fungal pathogen R. solani to reduce sclerotia formation. The experimental data revealed that the Inhibition rate (RH) for sclerotia formation was (15, 33, 74 and 93%), respectively, for their corresponding AuNPs concentrations (10, 20, 40 and 80 µg/mL). Our findings obviously indicated that the RH strongly depend on AuNPs rates, and enhance upon an increase in AuNPs rates. The application of endophytic fungi biomass for green synthesis is our future goal.

Comparison of three commercial DNA extraction kits for the enhancement of PCR assay sensitivity for Xanthomonas euvesicatoria pv. allii.

AIM: The study was carried out to evaluate three commercial DNA extraction kits, Probe GS, FitoSorb and Sorb GMO, thus identifying the most suitable for isolating the phytopathogenic bacteria Xanthomonas euvesicatoria pv. allii. MATERIALS AND METHODS: Onion seed samples were prepared which were inoculated with bacterial concentrations ranging from 101 to 107  CFU per ml. Real-time PCR was performed to determine the efficacy of the isolated DNA in enhancing the sensitivity of the assay. The DNA extracted by Probe GS had the best detectability, having been detected at the lowest concentration used in the study 101  × 3 CFU per ml. FitoSorb and Sorb GMO yielded DNA with a higher and similar limit of detection 103  × 3 CFU per ml. Furthermore, Probe GS had the lowest cycle at every concentration tested as compared to the other methods. CONCLUSION: Therefore, Probe GS proved to be the most optimized kit for the extraction of X. euvesicatoria pv. allii, hence enhanced degree of sensitivity for the assay. IMPACT AND SIGNIFICANCE OF THE STUDY: The findings generated in this study can be used by phytosanitary laboratories to develop highly rapid and accurate diagnostic protocols for X. euvesicatoria pv. allii.

Facile Biogenic Synthesis and Characterization of Seven Metal-Based Nanoparticles Conjugated with Phytochemical Bioactives Using Fragaria ananassa Leaf Extract.

In this investigation, for the first time, we used Fragaria ananassa (strawberry) leaf extract as a source of natural reducing, capping or stabilizing agents to develop an eco-friendly, cost-effective and safe process for the biosynthesis of metal-based nanoparticles including silver, copper, iron, zinc and magnesium oxide. Calcinated and non-calcinated zinc oxide nanoparticles also synthesized during a method different from our previous study. To confirm the successful formation of nanoparticles, different characterization techniques applied. UV-Vis spectroscopy, X-ray Diffraction (XRD) spectroscopy, Field Emission Scanning Electron Microscopy (FESEM) coupled with Energy Dispersive X-ray Spectroscopy (EDS), Photon Cross-Correlation Spectroscopy (PCCS) and Fourier Transformed Infrared Spectroscopy (FT-IR) were used to study the unique structure and properties of biosynthesized nanoparticles. The results show the successful formation of metal-based particles in the range of nanometer, confirmed by different characterization techniques. Finally, the presented approach has been demonstrated to be effective in the biosynthesis of metal and metal oxide nanoparticles.

How to Improve Sensitivity of Sandwich Lateral Flow Immunoassay for Corpuscular Antigens on the Example of Potato Virus Y?

A simple approach was proposed to decrease the detection limit of sandwich lateral flow immunoassay (LFIA) by changing the conditions for binding between a polyvalent antigen and a conjugate of gold nanoparticles (GNPs) with antibodies. In this study, the potato virus Y (PVY) was used as the polyvalent antigen, which affects economically important plants in the Solanaceae family. The obtained polyclonal antibodies that are specific to PVY were characterized using a sandwich enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). For LFIA, the antibodies were conjugated with GNPs with a diameter of 17.4 ± 1.0 nm. We conducted LFIAs using GNP conjugates in a dried state on the test strip and after pre-incubation with a sample. Pre-incubating the GNP conjugates and sample for 30 s was found to decrease the detection limit by 60-fold from 330 ng∙mL-1 to 5.4 ng∙mL-1 in comparison with conventional LFIA. The developed method was successfully tested for its ability to detect PVY in infected and uninfected potato leaves. The quantitative results of the proposed LFIA with pre-incubation were confirmed by ELISA, and resulted in a correlation coefficient of 0.891. The proposed approach is rapid, simple, and preserves the main advantages of LFIA as a non-laboratory diagnostic method.