Antifungal Activities of Pure and ZnO-Encapsulated Essential Oil of Zataria multiflora on Alternaria solani as the Pathogenic Agent of Tomato Early Blight Disease.

The utilization of plant essential oils (EOs) and nanomaterials due to their safety compared with synthetic chemicals has been considered in the management of plant diseases. In this study, the inhibitory effects of Zataria multiflora, Nepeta haussknechtii, Artemisia sieberi, and Citrus aurantifolia EOs in pure and Zinc Oxide (ZnO) nanocapsulated formulations were evaluated on the mycelial growth of Alternaria solani to find a suitable alternative for synthetic chemicals. The crystal structure and morphological properties of the fabricated nanomaterials were assessed via X-ray diffraction (XRD) and scanning electron microscope (SEM) analyses. The textural features of the prepared nanoparticles were investigated with Brunauer-Emmett-Teller (BET) analysis, and the presence of elements in the samples was studied with energy-dispersive X-ray (EDX) technique. The mycelial growth inhibitory (MGI) was performed in the laboratory by mixing with potato dextrose agar (PDA) medium at concentrations of 100, 300, 600, 1,000, 1,500, and 2,000 ppm. Based on the results, major differences were monitored between different concentrations. At the highest studied concentration, the inhibition of Z. multiflora EO was 100%, which was 43.20, 42.37, and 21.19% for N. haussknechtii, A. sieberi, and C. aurantifolia, respectively, and the inhibition of their nanocapsules was 100, 51.32, 55.23, and 26.58%, respectively. In the greenhouse study, Z. multiflora EO and its nanocapsule (ZnO-ZmEO) were compared with the ZnO and chlorothalonil fungicide based on the highest inhibitory of Z. multiflora in vitro. The highest antifungal effect was related to the ZnO-ZmEO by 53.33%. Therefore, the ZnO-ZmEO formulation can be recommended as a biofungicide for managing and controlling tomato early blight disease after further research.

Mycelial inhibitory effects of antagonistic fungi, plant essential oils and propolis against five phytopathogenic Fusarium species.

Fusarium spp. are considered as one of the most devastating plant pathogenic fungi worldwide. In this study, the effect of essential oil (EO) of Mentha longifolia, M. spicata, Achillea sp. and Foeniculum vulgare, ethanolic extract of Propolis (EEP), and Trichoderma harzianum T447 and T. hamatum T622 was investigated against five phytopathogenic Fusarium species. The results showed that the fungal species, the type of EO, and concentrations play a substantial role in inhibiting the mycelial growth of Fusarium spp. GC-MS analysis of the EOs showed that the piperitone oxide and cis-piperitone oxide were found as the main components of M. longifolia. Our results also revealed that EEP possessed the growth inhibitory effect against Fusarium spp. It was observed that the extracellular secretions of T. harzianum T447 showed very high inhibition against the fungi. Our results highlighted the need for further research to apply them as a safe alternative to the chemical pesticides.

Production of Innovative Essential Oil-Based Emulsion Coatings for Fungal Growth Control on Postharvest Fruits.

This work assessed the antimicrobial potential of natural essential oils (EOs) from cinnamon (CEO), zataria (ZEO), and satureja (SEO), applied natively or as coatings against Penicillium expansum and Botrytis cinerea during both in vitro and in vivo (on apple fruits) experiments. The induced inhibitory effect towards fungal growth, as a function of both EO type and concentration (75-1200 µL/L), was preliminarily investigated to select the most suitable EO for producing bacterial cellulose nanocrystals (BCNCs)/fish gelatin (GelA)-based emulsions. CEO and ZEO exhibited the best performances against P. expansum and B. cinerea, respectively. None of the pristine EOs completely inhibited the fungal growth and "disease severity", properly quantified via size measurements of lesions formed on fruit surfaces. As compared to pristine CEO, coating emulsions with variable CEO concentration (75-2400 µL/L) curbed lesion spreading on apples, owing to the controlled CEO release during a 21-day temporal window. The strongest effect was displayed by BCNCs/GelA-CEO emulsions at the highest CEO concentration, upon which lesions on fruit skins were barely detectable. This work demonstrated the capability of EOs embedded in BCNCs/GelA-based nanocapsules to efficiently slow down microbial spoilage on postharvest fruits, thus offering viable opportunities for developing innovative antimicrobial packaging systems.

Antifungal activity of TiO2/AgBr nanocomposites on some phytopathogenic fungi.

TiO2/AgBr composites were synthesized by a simple ultrasonic strategy. Various instruments such as SEM, EDX, XRD, and FT-IR were exploited to investigate their characteristics. Antifungal activities of the as-obtained samples were assessed through the inactivation of Fusarium graminearum in the spore suspension method and mycelial growth inhibition of F. graminearum, Botrytis cinerea, and Sclerotinia sclerotiorum in the microdilution method. The results represented that the TiO2/AgBr samples possess higher antifungal activities on F. graminearum spores than the pure TiO2. The sample with 20 wt% silver bromide represented the highest inhibitory effect on the growth of F. graminearum so that all fungal spores were degraded in the initial times of the treatment process. The inactivation of fungal spores after 60 min was 35.2%, 97.8%, 98.9%, and 98.7%, in respect, for 5, 10, 20, and 30 weight percent of AgBr in the binary nanocomposites, while the inhibition rate was 13.4% for the pure TiO2. With increasing ultrasound irradiation time for more than 30 min, the inactivation rate constant decreased. It was also found that the antifungal activity of the nanocomposites without calcination was higher than those of the calcined materials. Considering the antifungal potential against phytopathogenic fungi and advantages such as simple synthesis and eco-friendly nature, it seems that TiO2/AgBr nanocomposites can be used instead of synthetic chemicals after additional field investigations and mass production.

Control of nectarine fruits postharvest fungal rots caused by Botrytis Cinerea and Rhizopus Stolonifer via some essential oils.

Nectarines (Prunus persica L. Bath) are very sensitive fruit to fungal infection. Today, the control of postharvest fruit diseases with essential oils (EOs) has been significantly noticed as a novel trend in biological preservation. In this study, volatile compounds of Cinnamon zeylanicum (CEO), Zataria multiflora (ZEO), and Satureja khuzestanica (SEO) were analyzed by Gas Chromatography-Mass spectroscopy. Also, the in vitro antifungal activities of EOs against Botrytis cinerea and Rhizopus stolonifer were evaluated at different concentrations. The in vivo antifungal activity of these EOs on artificially infected nectarine fruits was also considered. The major components were Thymol (32.68%) and Carvacrol (30.57%) for ZEO, cinnamaldehyde (80.82%) for CEO, and carvacrol (38.43%) for SEO. The application of different concentrations showed a decreasing trend in the fungus radial growth in all EOs. In the in vitro experiments, ZEO and CEO exhibited more significant mycelial inhibition results and reduction of the IC50, MIC and MFC values against Botrytis cinerea and Rhizopus stolonifer, respectively. However, in the in vitro experiments, none of the treatments were capable of completely inhibiting the growth of the fungi. According to the results of this study, ZEO and CEO could reduce the damage caused by these fungi.

Separate and Combined Effects of Mentha piperata and Mentha pulegium Essential Oils and a Pathogenic Fungus Lecanicillium muscarium Against Aphis gossypii (Hemiptera: Aphididae).

In the present study, the toxicity of essential oils of Mentha piperata L. and Mentha pulegium L. and pathogenicity of Lecanicillium muscarium (Zare & Gams) were studied in the melon aphid, Aphis gossypii Glover. Analyses of the essential oils by GC-MS indicated limonene (27.28%), menthol (24.71%), menthone (14.01%), and carvol (8.46%) in the M. piperata essential oil and pulegone (73.44%), piperitenone (5.49%), decane (4.99%), and limonene (3.07%) in the essential oil of M. pulegium as the main components. Both essential oils and the pathogenic fungus had useful toxicity against A. gossypii. Probit analysis indicated LC50 values (lethal concentrations to kill 50% of population; 95% confidence limits in parentheses) of M. piperata and M. pulegium essential oils as 15.25 (12.25-19.56) and 23.13 (19.27-28.42) µl/liter air, respectively. Susceptibility to the pathogenic fungus increased with exposure time. Aphid mortality also increased when the essential oils were combined with L. muscarium, although the phenomena was additive rather than synergistic. Mycelial growth inhibition of L. muscarium exposed to the essential oils was also very low. Based on our results, M. piperata and M. pulegium essential oils and the pathogenic fungus L. muscarium have some potential for management of A. gossypii.

Rapid identification of Fusarium graminearum species complex using Rolling Circle Amplification (RCA).

Rolling Circle Amplification (RCA) of DNA is a sensitive and cost effective method for the rapid identification of pathogenic fungi without the need for sequencing. Amplification products can be visualized on 1% agarose gel to verify the specificity of probe-template binding or directly by adding fluorescent dyes. Fusarium Head Blight (FHB) is currently the world's largest threat to the production of cereal crops with the production of a range of mycotoxins as an additional risk. We designed sets of RCA padlock probes based on polymorphisms in the elongation factor 1-α (EF-1α) gene to detect the dominant FHB species, comprising lineages of the Fusarium graminearum species complex (FGSC). The method also enabled the identification of species of the Fusarium oxysporum (FOSC), the Fusarium incarnatum-equiseti (FIESC), and the Fusarium tricinctum (FTSC) species complexes, and used strains from the CBS culture collection as reference. Subsequently probes were applied to characterize isolates from wheat and wild grasses, and inoculated wheat kernels. The RCA assays successfully amplified DNA of the target fungi, both in environmental samples and in the contaminated wheat samples, while no cross reactivity was observed with uncontaminated wheat or related Fusarium species. As RCA does not require expensive instrumentation, the technique has a good potential for local and point of care screening for toxigenic Fusarium species in cereals.

Theoretical investigations on the hydrolysis pathway of tin verdoheme complexes: elucidation of tin's ring opening inhibition role.

In order to obtain a better molecular understanding of inhibitory role of tin metal in the verdoheme ring opening process, hydrolysis of three possibly six, five, and four coordinate verdoheme complexes of tin(IV) and (II) have been studied using DFT method. The results of calculations indicate that, in excellent accord with experimental reports, hydrolysis of different possibly coordinated tin(IV) and (II) verdohemes does not lead to the opening of the macrocycle. Contrary to iron and zinc verdohemes, in five and four coordinate verdoheme complexes of tin(IV) and (II), formation of open ring helical complexes of tin are unfavorable both thermodynamically and kinetically. In these pathways, coordination of hydroxide nucleophile to tin metal due to the highly charged, exclusive oxophilicity nature of the Sn center, and high affinity of Sn to increase coordination state are proposed responsible as inhibiting roles of tin via the ring opening. While, in saturated six coordinate tin(IV) and (II) verdoheme complexes the ring opening of tin verdohemes is possible thermodynamically, but it is not predicted to occur from a kinetics point of view. In the six coordinate pathway, tin plays no coordination role and direct addition of hydroxide nucleophile to the positive oxo-carbon centers and formation of closed ring hydroxy compounds is proposed for preventing the verdoheme ring opening. These key points and findings have been corroborated by the results obtained from atomic charge analysis, geometrical parameters, and molecular orbital calculations. In addition, the results of inhibiting ring opening reaction of tin verdoheme complexes could support the great interest of tin porphyrin analogues as pharmacologic means of chemoprevention of neonatal jaundice by the competitive inhibitory action of tin porphyrins on heme oxygenase.