Attenuation of Zucchini mosaic virus disease in cucumber plants by mycorrhizal symbiosis.

KEY MESSAGE: Arbuscular mycorrhizal fungi generated systemic acquired resistance in cucumber to Zucchini yellow mosaic virus, indicating their prospective application in the soil as a sustainable, environmentally friendly approach to inhibit the spread of pathogens. The wide spread of plant pathogens affects the whole world, causing several plant diseases and threatening national food security as it disrupts the quantity and quality of economically important crops. Recently, environmentally acceptable mitigating practices have been required for sustainable agriculture, restricting the use of chemical fertilizers in agricultural areas. Herein, the biological control of Zucchini yellow mosaic virus (ZYMV) in cucumber (Cucumis sativus L.) plants using arbuscular mycorrhizal (AM) fungi was investigated. Compared to control plants, ZYMV-infected plants displayed high disease incidence (DI) and severity (DS) with various symptoms, including severe yellow mosaic, mottling and green blisters of leaves. However, AM fungal inoculation exhibited 50% inhibition for these symptoms and limited DS to 26% as compared to non-colonized ones. The detection of ZYMV by the Enzyme-Linked Immunosorbent Assay technique exhibited a significant reduction in AM-inoculated plants (5.23-fold) compared with non-colonized ones. Besides, mycorrhizal root colonization (F%) was slightly reduced by ZYMV infection. ZYMV infection decreased all growth parameters and pigment fractions and increased the malondialdehyde (MDA) content, however, these parameters were significantly enhanced and the MDA content was decreased by AM fungal colonization. Also, the protein, proline and antioxidant enzymes (POX and CAT) were increased with ZYMV infection with more enhancements due to AM root colonization. Remarkably, defence pathogenesis-related (PR) genes such as PR-a, PR-b, and PR-10 were quickly expressed in response to AM treatment. Our findings demonstrated the beneficial function of AM fungi in triggering the plant defence against ZYMV as they caused systemic acquired resistance in cucumber plants and supported their potential use in the soil as an environment-friendly method of hindering the spread of pathogenic microorganisms sustainably.

Trichoderma cf. asperellum and plant-based titanium dioxide nanoparticles initiate morphological and biochemical modifications in Hordeum vulgare L. against Bipolaris sorokiniana.

BACKGROUND: Spot blotch is a serious foliar disease of barley (Hordeum vulgare L.) plants caused by Bipolaris sorokiniana, which is a hemibiotrophic ascomycete that has a global impact on productivity. Some Trichoderma spp. is a promising candidate as a biocontrol agent as well as a plant growth stimulant. Also, the application of nanomaterials in agriculture limits the use of harmful agrochemicals and helps improve the yield of different crops. The current study was carried out to evaluate the effectiveness of Trichoderma. cf. asperellum and the biosynthesized titanium dioxide nanoparticles (TiO2 NPs) to manage the spot blotch disease of barley caused by B. sorokiniana and to assess the plant's innate defense response. RESULTS: Aloe vera L. aqueous leaf extract was used to biosynthesize TiO2 NPs by reducing TiCl4 salt into TiO2 NPs and the biosynthesized NPs were detected using SEM and TEM. It was confirmed that the NPs are anatase-crystalline phases and exist in sizes ranging from 10 to 25 nm. The T. cf. asperellum fungus was detected using morphological traits and rDNA ITS analysis. This fungus showed strong antagonistic activity against B. sorokiniana (57.07%). Additionally, T. cf. asperellum cultures that were 5 days old demonstrated the best antagonistic activity against the pathogen in cell-free culture filtrate. Also, B. sorokiniana was unable to grow on PDA supplemented with 25 and 50 mg/L of TiO2 NPs, and the diameter of the inhibitory zone increased with increasing TiO2 NPs concentration. In an in vivo assay, barley plants treated with T. cf. asperellum or TiO2 NPs were used to evaluate their biocontrol efficiency against B. sorokiniana, in which T. cf. asperellum and TiO2 NPs enhanced the growth of the plant without displaying disease symptoms. Furthermore, the physiological and biochemical parameters of barley plants treated with T. cf. asperellum or TiO2 NPs in response to B. sorokiniana treatment were quantitively estimated. Hence, T. cf. asperellum and TiO2 NPs improve the plant's tolerance and reduce the growth inhibitory effect of B. sorokiniana. CONCLUSION: Subsequently, T. cf. asperellum and TiO2 NPs were able to protect barley plants against B. sorokiniana via enhancement of chlorophyll content, improvement of plant health, and induction of the barley innate defense system. The present work emphasizes the major contribution of T. cf. asperellum and the biosynthesized TiO2 NPs to the management of spot blotch disease in barley plants, and ultimately to the enhancement of barley plant quality and productivity.

Unique properties of titanium dioxide quantum dots assisted regulation of growth and biochemical parameters of Hibiscus sabdariffa plants.

Owing to the uniqueness of quantum dots (QDs) as a potential nanomaterial for agricultural application, hence in the present study, titanium dioxide quantum dots (TiO2 QDs) were successfully synthesized via sol-gel technique and the physico-chemical properties of the prepared TiO2 QDs were analyzed. Based on the results, the TiO2 QDs showed the presence of anatase phase of TiO2. TEM examination revealed spherical QDs morphology with an average size of 7.69 ± 1.22 nm. The large zeta potential value (-20.9 ± 2.3 mV) indicate greater stability of the prepared TiO2 QDs in aqueous solutions. Moreover, in this work, the application of TiO2 QDs on Hibiscus sabdariffa plants was conducted, where H. sabdariffa plants were foliar sprayed twice a week in the early morning with different concentrations of TiO2 QDs (0, 2, 5, 10, 15 and 30 ppm) to evaluate their influence on these plants in terms of morphological indexes and biochemical parameters. The results exhibited an increasing impact of the different used concentrations of TiO2 QDs on morphological indexes, such as fresh weight, dry weight, shoot length, root length, and leaf number, and physio-biochemical parameters like chlorophyll a, chlorophyll b, carotenoid contents, total pigments and total phenolic contents. Remarkably, the most prominent result was recorded at 15 ppm TiO2 QDs where plant height, total protein and enzymatic antioxidants like catalase and peroxidase were noted to increase by 47.6, 20.5, 29.5 and 38.3%, respectively compared to control. Therefore, foliar spraying with TiO2 QDs positively serves as an effective strategy for inducing optimistic effects in H. sabdariffa plants.

In vivo and In vitro evaluation of the antifungal activity of the PGPR Bacillus amyloliquefaciens RaSh1 (MZ945930) against Alternaria alternata with growth promotion influences on Capsicum annuum L. plants.

Alternaria alternata that threatens pepper production and causes major economic harm is responsible for the leaf spot/blight disease. Chemical fungicides have been widely employed; unfortunately, fungicidal resistance is a current concern. Therefore, finding new environmentally friendly biocontrol agents is a future challenge. One of these friendly solutions is the use of bacterial endophytes that have been identified as a source of bioactive compounds. The current study investigates the in vivo and in vitro fungicidal potential of Bacillus amyloliquefaciens RaSh1 (MZ945930) against pathogenic A. alternata. In vitro, the results revealed that RaSh1 exhibited strong antagonistic activity against A. alternata. In addition to this, we inoculated pepper (Capsicum annuum L.) plants with B. amyloliquefaciens RaSh1 and infected them with A. alternata. As a result of A. alternata infection, which generated the highest leaf spot disease incidence (DI), the plant's growth indices and physio-biochemical characteristics significantly decreased, according to our findings. Our results also showed the abnormal and deformed cell structure using light and electron microscopy of A. alternata-infected leaves compared with other treatments. However, DI was greatly reduced with B. amyloliquefaciens RaSh1 application (40%) compared to pepper plants infected with A. alternata (80%), and this led to the largest increases in all identified physio-biochemical parameters, including the activity of the defense-related enzymes. Moreover, inoculation of pepper plants with B. amyloliquefaciens RaSh1 decreased electrolyte leakage by 19.53% and MDA content by 38.60% as compared to A. alternata infected ones. Our results show that the endophyte B. amyloliquefaciens RaSh1 has excellent potential as a biocontrol agent and positively affects pepper plant growth.

Assessment of beneficial fungal microorganism's bio-efficacy in stimulating morphological and physiological parameters of Allium cepa plants grown in soil amended with fish wastes.

BACKGROUND: The increase in the human consumption of fish results in the production of organic fish wastes (FW). For enhanced soil fertility and plant growth at a lower cost and without the negative impacts of chemical fertilizers, these wastes could be employed as a valuable organic fertilizer. To determine the synergistic bio-efficacy of Trichoderma sp. and arbuscular mycorrhizal (AM) fungi in stimulating the morphological and physiological characteristics of FW-fertilized Alium cepa, as well as to investigate their involvement in boosting soil fertility, the current study was carried out. Overall, eight treatments were applied as follows: AM, Trichoderma sp., AM + Trichoderma sp., FW, AM + FW, Trichoderma sp. + FW, AM + Trichoderma sp. + FW, and control. Growth and physiological assessments of onion plants were taken after 8 weeks from FW application. RESULTS: Our results showed that FW application combined with AM fungi and Trichoderma sp. inoculations increased aggregate stability of the soil (glomalin content) and soil chitinase activity. Moreover, using the bio-inoculations along with FW amendments significantly (p < 0.05) improved the photosynthetic pigments, protein, carbohydrates, and nutrients content of onion plants. It's interesting to note that the triple interaction of AM + Trichoderma sp. + FW led to the greatest increase in plant height, root length, number of leaves, and leaf area as well as total fresh and dry weights of shoots and roots. Besides, AM fungal colonization was at its highest percentage with Trichoderma sp. inoculation, although this percentage decreased with FW addition. CONCLUSION: We concluded that the combined treatments of AM fungi and Trichoderma sp. along with FW application to the soil can be proposed as a successful strategy for plant performance in nutrient-deficient soils as both fungal inoculants are capable of degrading these wastes and converting them into manure suitable for farming so plants can uptake the minerals effortlessly.

Potential of green synthesized titanium dioxide nanoparticles for enhancing seedling emergence, vigor and tolerance indices and DPPH free radical scavenging in two varieties of soybean under salinity stress.

BACKGROUND: Considering titanium dioxide nanoparticles (TiO2 NPs) role in plant growth and especially in plant tolerance against abiotic stress, in the present work, TiO2 NPs were green synthesized using an aqueous solution of Aloe vera leaf extract as a capping agent and titanium tetrachloride as a precursor. These green synthesized TiO2 NPs were characterized using different techniques: UV spectrophotometer, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD). Results revealed that synthesized TiO2 NPs possess a tetragonal morphology with a size ranging from 10 to 25 nm. Additionally, the present work evaluated the effects of three concentrations of TiO2 NPs (0, 30 and 50 ppm) and six NaCl concentrations (0, 25, 50, 100, 150 and 200 mM) and their interactions with respect to germination parameters, vigor indices, oxidative stress and DPPH free radical scavenging of two varieties of soybean (Glycine max L. var. 22 and 35). RESULTS: Results demonstrated that all germination traits and vigor indices were negatively affected under all salinity levels. Also, the contents of hydrogen peroxide (H2O2) and malondialdehyde (MDA) were significantly increased by increasing the NaCl concentrations in two soybean varieties. Most interestingly, TiO2 NPs (30 ppm) mediated positive effects on germination parameters, reducing H2O2 and MDA contents by enhancing antioxidant (decreasing IC50) whereas 50 ppm showed an intermediate response under both control and saline soil conditions. CONCLUSION: Our findings demonstrate the growth enhancement effects of TiO2 NPs application as well as its ameliorative potential in dealing with salinity.

Potential use of beneficial fungal microorganisms and C-phycocyanin extract for enhancing seed germination, seedling growth and biochemical traits of Solanum lycopersicum L.

BACKGROUND: Biopriming as a new technique of seed treatment involves the application of beneficial microorganisms on the seed surface to stimulate seed germination, plant growth, and protect the seed from soil and seed-borne pathogens. The present investigation was carried out on seed germination, seedling vigor and biochemical traits of one of the most important vegetable crops (Tomato, Solanum lycopersicum L.). The treatments comprised viz. T1: Non primed seeds (Control), T2: Hydropriming, T3: Biopriming with C-phycocyanin (C-PC) (Spirulina platensis extract), T4: Biopriming with Trichoderma asperellum, T5: Biopriming with T. viride, T6: Biopriming with Beauveria bassiana. RESULTS: Extraction and purification of C-phycocyanin (C-PC) from the dry S. platensis powder using various methods was performed. The purity after dialyses was 0.49 and its ultimate purity (A620/A280) after ion-exchange chromatography was 4.64. The results on tomato seedlings revealed that the maximum germination percentage (100%), germination index (15.46 and 15.12), seedling length (10.67 cm), seedling dry weight (1.73 and 1.97 mg) and seedling length vigor index (1066.7) were recorded for tomato biopriming with T. viride, and B. bassiana (T5 and T6). Moreover, the quantitative estimation of total carbohydrates and total free amino acids contents in bioprimed tomato seedlings indicated a significantly higher amount with T. viride, followed by those bioprimed with T. asperellum, B. bassiana and C-PC extract. CONCLUSION: Thus, our results indicated that biopriming of tomato seeds with beneficial fungal inoculants and C-PC was very effective. The most operative biostimulants were those bioprimed with T. viride and B. bassiana compared to other biostimulants (T. asperellum and C-PC). Therefore, to ensure sustainable agriculture, this study offers new possibilities for the biopriming application as an alternative and ecological management strategy to chemical treatment and provides a valuable basis for improving seed germination.

Physiological Responses of Salinized Fenugreek (Trigonellafoenum-graecum L.) Plants to Foliar Application of Salicylic Acid.

Considering the detrimental effects of salt stress on the physiological mechanisms of plants in terms of growth, development and productivity, intensive efforts are underway to improve plant tolerance to salinity. Hence, an experiment was conducted to assess the impact of the foliar application of salicylic acid (SA; 0.5 mM) on the physiological traits of fenugreek (Trigonellafoenum-graecum L.) plants grown under three salt concentrations (0, 75, and 150 mM NaCl). An increase in salt concentration generated a decrease in the chlorophyll content index (CCI); however, the foliar application of SA boosted the CCI. The malondialdehyde content increased in salt-stressed fenugreek plants, while a reduction in content was observed with SA. Likewise, SA application induced an accumulation of proline, total phenolics, and flavonoids. Moreover, further increases in total free amino acids and shikimic acid were observed with the foliar application of SA, in either control or salt-treated plants. Similar results were obtained for ascorbate peroxidase, peroxidase, polyphenol oxidase, and catalase with SA application. Hence, we concluded that the foliar application of SA ameliorates salinity, and it is a growth regulator that improves the tolerance of fenugreek plants under salt stress.

Tracking of Zinc Ferrite Nanoparticle Effects on Pea (Pisum sativum L.) Plant Growth, Pigments, Mineral Content and Arbuscular Mycorrhizal Colonization.

Important gaps in knowledge remain regarding the potential of nanoparticles (NPs) for plants, particularly the existence of helpful microorganisms, for instance, arbuscular mycorrhizal (AM) fungi present in the soil. Hence, more profound studies are required to distinguish the impact of NPs on plant growth inoculated with AM fungi and their role in NP uptake to develop smart nanotechnology implementations in crop improvement. Zinc ferrite (ZnFe2O4) NPs are prepared via the citrate technique and defined by X-ray diffraction (XRD) as well as transmission electron microscopy for several physical properties. The analysis of the XRD pattern confirmed the creation of a nanocrystalline structure with a crystallite size equal to 25.4 nm. The effects of ZnFe2O4 NP on AM fungi, growth and pigment content as well as nutrient uptake of pea (Pisum sativum) plants were assessed. ZnFe2O4 NP application caused a slight decrease in root colonization. However, its application showed an augmentation of 74.36% and 91.89% in AM pea plant shoots and roots' fresh weights, respectively, compared to the control. Moreover, the synthesized ZnFe2O4 NP uptake by plant roots and their contents were enhanced by AM fungi. These findings suggest the safe use of ZnFe2O4 NPs in nano-agricultural applications for plant development with AM fungi.

The individual and interactive role of arbuscular mycorrhizal fungi and Trichoderma viride on growth, protein content, amino acids fractionation, and phosphatases enzyme activities of onion plants amended with fish waste.

The Green Revolution faced a great cost to meet ever-increasing demands for food, where indiscriminate use of agrochemicals resulted in non-friendly habitats. Therefore, the development of a sustainable approach to better crop production of onion seeds (Allium cepa L.) is very crucial. It is time to use organic waste as a replacement for agrochemicals by using arbuscular mycorrhizal fungi (AMF) and Trichoderma. Fish waste as representative of food waste acts as a leading cause of contamination of the environment. The interaction of AMF and Trichoderma viride on biomass, total soluble protein, mycorrhizal colonization, amino acids, phosphatases and phosphorus and nitrogen contents of onion plants grown in fish waste amended soil was studied. Fish waste has caused a slight increase in onions biomass, total free amino acids, and soluble protein content while with AMF and T. viride dual inoculation more increments were recorded; such increases were related to an increase in mycorrhizal colonization. T. viride application significantly increased the mycorrhizal colonization levels, but these were significantly reduced with waste addition. Analysis of amino acids in plants showed that their concentrations had changed as a result of waste addition combined with AMF and/or T. viride. The effectiveness of fish waste combined with low cost and health/environmental safety leads to a prediction that the introduction of fish waste coupled with fungi will become a more popular feature of agriculture in the future.

Alleviation of cadmium stress by arbuscular mycorrhizal symbiosis.

Owing to the realization of the harmful effect of cadmium on the environment and plants and as the plants are sessile organisms, they need to increase the protective mechanisms to cope with Cd stress. Inoculation the plant with soil microbes at the place of their growing is an important strategy to support the plants against stresses. In this study, trigonella plants were inoculated with arbuscular mycorrhizal (AM) fungi under different CdCl2 concentrations (0, 2.25, and 6.25 mM). AM inoculation increased growth parameters, chlorophyll, and protein contents. Root colonization was significantly increased at low Cd concentration (2.25 mM) and decreased at high one (6.25 mM). Also, with AM fungal inoculation, the translocation factor of trigonella plants significantly decreased as compared to non-AM ones at both low and high Cd concentrations. In addition, it was clearly that malondialdehyde content of trigonella plants increased significantly at both Cd concentrations and with AM fungal inoculation its content decreased compared to those of non-AM ones. AM inoculation significantly increased antioxidant enzymes activities compared to non-AM ones. Consequently, this study showed a tolerance strategy of AM trigonella plants against Cd stress, thus mycorrhizal symbiosis becomes a promising and suitable as phytostabilizers of Cd stressed soil.