Green synthesis of silver nanoparticles by sweet cherry and its application against cherry spot disease.

Asia has a rich history of cultivating sweet cherries, a practice that has been carried out since ancient times. However, the effective management of Alternaria disease in sweet cherry crops has presented a formidable challenge, resulting in notable decreases in yield. Various attempts have been made to employ both chemical and biological treatments; however, their effectiveness has been restricted. In order to tackle this problem, an investigation was carried out, with the primary objective of isolating and identifying Alternaria isolates that are accountable for the occurrence of sweet cherry soft spot rot. Out of the twelve isolates examined, the CHM-4 isolate was found to be the most pathogenic. Its identification was achieved through the use of the ITS genomic region (ITS1 and ITS4), and the BLAST results revealed a 95 % similarity with Alternaria alternata (MG744381.1). The objective of the research was to explore the potential of silver nanoparticles (SNPs) synthesized by phytosynthesis as a novel antifungal agent to combat sweet cherry soft spot pathogenicity. The biosynthesis of SNPs was carried out using sweet cherry fruits kernel exudate, which served as an environmentally friendly source. The exudates exhibited the ability to produce nanoparticles with an average size of 24.97 nm. Analysis conducted using a transmission electron microscope (TEM) revealed the multifaceted structure of these nanoparticles. Furthermore, when tested at concentrations of 5, 10, 20, and 40 µg/ml, these biosynthetic nanoparticles demonstrated the capability to inhibit the growth of Alternaria fungi and effectively destroy fungal hyphae. It is advisable to utilize diverse components of sweet cherry for the synthesis of various nanoparticles owing to their compatibility with the surrounding environment.

Impact of different types of green manure on pests and disease incidence and severity as well as growth and yield parameters of maize.

The emergence of pests and diseases, including the maize streak virus, leaf blight, the African stem borer, and gray leaf spot, poses a persistent threat to maize production (Zea mays L. cv: DMR-ESR-Yellow) around the world. A field experiment was conducted at the School of Agriculture experimental site, Njala University, Sierra Leone, during a two-year period (2020-2021) to assess the effects of green manure on pest and disease incidence and severity as well as growth and yield parameters of maize. The experiment was laid out in a randomized complete block design (RCBD) with three replications and four treatments: Cal. 3 t.ha-1, Cal. 6 t.h-1, Pan. 3 t.h-1, Pan 6 t.ha-1 and a control plot amended with 200 kg ha-1 of N (urea) and NPK 15:15:15 ha-1 split application. The study showed that gray leaf spot damage was the most severe infection among all treatments. Therefore, the effects of the most severe disease and pest of maize in Sierra Leone can be minimized by applying green manure. Moreover, results reveal that Calopogonium- Pueraria mixture amended plots showed significant performance in the measured growth parameters viz. highest leaf number, large leaf area stem girth, superior plant height, best ear height (64.6-78.5 cm), higher cob yield (1.2-1.4 t.ha-1) ear (1.8-2.1 t.ha-1) and dry grain yield (0.5-0.7 ha-1). Panicum green manure results showed that prompt and adequate application, as well as decomposition of green manures, is imperative for the successful conservation and sustainability of maize farming systems. The findings of this research could improve the efficiency of green manure use in pest, disease, and crop management systems.

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.

In Vitro Evaluation of Antibacterial and Antifungal Activity of Biogenic Silver and Copper Nanoparticles: The First Report of Applying Biogenic Nanoparticles against Pilidium concavum and Pestalotia sp. Fungi.

There is increased attention paid to metallic nanoparticles due to their intensive use in various branches of agriculture and biotechnology, such as pest management, nanosensors, gene delivery, seed treatment, etc. There has been growing interest in applying environmentally friendly strategies for synthesizing nanoparticles without using substances which are hazardous to the environment. Biological practices for the synthesis of nanoparticles have been considered as possible ecofriendly alternatives to chemical synthesis. In the present study, we used biogenic silver and copper nanoparticles which were prepared by a previously reported green method. Moreover, the problem of chemical residues, which usually remain along with chemically synthesized nanoparticles and limit their application, was solved by developing such a green synthesis approach. To study the antibacterial activity of silver and copper nanoparticles, Pseudomonas aeruginosa was used; for the evaluation of antifungal activity, the pathogenic fungi Botrytis cinerea, Pilidium concavum and Pestalotia sp. were applied. To the best of our knowledge, this study represents the first time that the antifungal impact of a nanoparticle has been tested on Pilidium concavum and Pestalotia sp. Silver nanoparticles were found to be the more effective antimicrobial agent against all examined pathogens in comparison to copper nanoparticles. Data from such investigations provide valuable preliminary data on silver nanoparticle-based compounds or composites for use in the management of different pathogens.

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.

Impact of foliar fertilization on apple and pear trees in reconciling productivity and alleviation of environmental concerns under arid conditions.

Drought and heat stress are significant factors limiting fruit crop yield in arid conditions. Foliar fertilization is a common practice of supplying fruit crop production with mineral nutrients, especially under limited soil nutrient availability conditions. To evaluate potential effectiveness of the foliar application of macro-, micronutrient and growth regulators on dynamics of physiological parameters of the pear and apple cultivars under abiotic stresses, three-year experiments were carried out under arid conditions at the Russian Research Institute of Arid Agriculture during the 2015-2017 growing seasons. It has been revealed that foliar nutrition reduces the negative influence of heat stress, stabilizes the functional state of plants, thereby enhancing resistance to drought. During the most severe drought periods of vegetation, under the influence of foliar nutrition, there was a significant increase in the total water content (TWC), relative leaf turgidity (RLT) and water retention capacity (WRC); and also index of leaf water deficiency (LWD) was improved as compared to the non-treated control. All foliar treatments involving the macro-, micronutrient and growth regulators significantly enhanced fruit crop yield of pear and apple varieties over the control, yield enhancement was obtained 2.7-22.0 t ha-1 for the Talgar beauty (pear variety), 2.2-19.3 t ha-1 for the Renet Symirenko (apple variety), and 1.6-10.5 t ha-1 for the Starkrimson (apple variety). The most effective treatments for water consumption coefficient (WCC) were plantafol and speedphol. The results suggest that foliar plantafol and speedfol could be used as part of an efficient, sustainable fertilizer program for apple and pear trees for maintaining or improving fruit quality, productivity, and avoiding negative efficacy of abiotic stresses.

Enlargement of Gold Nanoparticles for Sensitive Immunochromatographic Diagnostics of Potato Brown Rot.

Lateral flow immunoassay (LFIA) is a convenient tool for rapid field-based control of various bacterial targets. However, for many applications, the detection limits obtained by LFIA are not sufficient. In this paper, we propose enlarging gold nanoparticles' (GNPs) size to develop a sensitive lateral flow immunoassay to detect Ralstonia solanacearum. This bacterium is a quarantine organism that causes potato brown rot. We fabricated lateral flow test strips using gold nanoparticles (17.4 ± 1.0 nm) as a label and their conjugates with antibodies specific to R. solanacearum. We proposed a signal enhancement in the test strips' test zone due to the tetrachloroauric (III) anion reduction on the GNP surface, and the increase in size of the gold nanoparticles on the test strips was approximately up to 100 nm, as confirmed by scanning electron microscopy. Overall, the gold enhancement approach decreased the detection limit of R. solanacearum by 33 times, to as low as 3 × 104 cells∙mL⁻1 in the potato tuber extract. The achieved detection limit allows the diagnosis of latent infection in potato tubers. The developed approach based on gold enhancement does not complicate analyses and requires only 3 min. The developed assay together with the sample preparation and gold enlargement requires 15 min. Thus, the developed approach is promising for the development of lateral flow test strips and their subsequent introduction into diagnostic practice.

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.