Protective role of iron oxide nanocomposites on disease index, and biochemical resistance indicators against Fusarium oxysporum induced-cucumber wilt disease: In vitro, and in vivo studies.

Recently nanocomposites have become a super-growth inducers as well as vital antifungal agents, which enhance plant growth and suppress plant diseases. A new strategy regarding the fabrication of humic acid (H) and boron (B) conjugated Fe2O3 nanocomposites was performed. Fe2O3 NP-B and Fe2O3 NP-H were synthesized in the presence of gamma-rays (as a direct reducing agent). Gamma-rays provoked reduction of metal ions due to the liberated reducing electrons, (e-aq), in aqueous solutions which can be considered as a direct reduction. Antifungal potential against Fusarium oxysporum, the causative agent of wilt disease in cucumber was determined. Disease index percent, metabolic resistance indicators in cucumber plant as response to promotion of systemic resistance (SR) were recorded. Results illustrated that both Fe2O3 NPs-B and Fe2O3 NPs-H nanocomposites had antifungal activity against F. oxysporumin vitro as well as in vivo. Results revealed that minimum inhibitory concentrations of Fe2O3 NPs-B and Fe2O3 NPs-H nanocomposites were 0.25 and 0.125 mM, respectively. Application of Fe2O3 NPs-B (0.25 mM) and Fe2O3 NPs-H (0.125 mM) appeared highly reduced the cucumber wilt disease symptoms incidence caused by F. oxysporum, and recorded disease severity by 83.33%. Fe2O3 NPs-B was the best treatment reducing disease indexes by 20.83% and gave highly protection against wilt disease by 75.0% and came next Fe2O3 NPs-H which reduced disease indexes by 25% and gave 69.99% protection against disease. Fe2O3 NPs-B and Fe2O3 NPs-H treatments improved morphological traits, photosynthetic pigments, osmolytes, total phenol and antioxidant enzymes activities in both infected and non-infected plants. The beneficial effects of the synthesized Fe2O3 NPs-B and Fe2O3 NPs-H nanocomposites were extended to increase not only the total phenol, and total soluble protein contents but also the activities of peroxidase (POD), and polyphenol oxidase (PPO) enzymes of the healthy and infected cucumber plants in comparison with control.

Ziziphus spina-christi extract-stabilized novel silver nanoparticle synthesis for combating Fusarium oxysporum-causing pepper wilt disease: in vitro and in vivo studies.

The novelty of the present study is studying the ability of aqueous Ziziphus spina-christi leaves' extract (ZSCE) to produce eco-friendly and cost-effective silver nanoparticles (Ag NPs) against Fusarium wilt disease. Phytochemical screening of ZSCE by HPLC showed that they contain important antimicrobial substances such as Rutin, Naringin, Myricetin, Quercetin, Kaempferol, Hesperidin, Syringeic, Eugenol, Pyrogallol, Gallic and Ferulic. Characterization methods reveal a stable Ag NPs with a crystalline structure, spherical in shape with average particle size about 11.25 nm. ZSCE and Ag NPs showed antifungal potential against F. oxysporum at different concentrations with MIC of Ag NPs as 0.125 mM. Ag NPs treatment was the most effective, as it gave the least disease severity (20.8%) and the highest protection rate (75%). The application of ZSCE or Ag NPs showed a clear recovery, and its effectiveness was not limited for improving growth and metabolic characteristics only, but also inducing substances responsible for defense against pathogens and activating plant immunity (such as increasing phenols and strong expression of peroxidase and polyphenol oxidase as well as isozymes). Owing to beneficial properties such as antifungal activity, and the eco-friendly approach of cost and safety, they can be applied in agricultural field as novel therapeutic nutrients.

Bioactive compounds and biomedical applications of endophytic fungi: a recent review.

Human life has been significantly impacted by the creation and spread of novel species of antibiotic-resistant bacteria and virus strains that are difficult to manage. Scientists and researchers have recently been motivated to seek out alternatives and other sources of safe and ecologically friendly active chemicals that have a powerful and effective effect against a wide variety of pathogenic bacteria as a result of all these hazards and problems. In this review, endophytic fungi and their bioactive compounds and biomedical applications were discussed. Endophytes, a new category of microbial source that can produce a variety of biological components, have major values for study and broad prospects for development. Recently, endophytic fungi have received much attention as a source for new bioactive compounds. In addition, the variety of natural active compounds generated by endophytes is due to the close biological relationship between endophytes and their host plants. The bioactive compounds separated from endophytes are usually classified as steroids, xanthones, terpenoids, isocoumarins, phenols, tetralones, benzopyranones and enniatines. Moreover, this review discusses enhancement methods of secondary metabolites production by fungal endophytes which include optimization methods, co-culture method, chemical epigenetic modification and molecular-based approaches. Furthermore, this review deals with different medical applications of bioactive compounds such as antimicrobial, antiviral, antioxidant and anticancer activities in the last 3 years.

Potential of biosynthesized zinc oxide nanoparticles to control Fusarium wilt disease in eggplant (Solanum melongena) and promote plant growth.

In this study, a novel, non-toxic, eco-friendly zinc oxide nanoparticles (ZnO-NPs) was used instead of the synthetic fungicides widely used to control the destructive phytopathogenic fungus Fusarium oxysporum, the causative agent of wilt disease in Solanum melongena L. Herein, the biosynthesized ZnO-NPs was carried out by Penicillium expansum ATCC 7861. In vitro, mycosynthesized ZnO-NPs exhibited antifungal activity against Fusarium oxysporum. In vivo, ZnO-NPs suppressed Fusarium wilt disease in cultivated Solanum melongena L. by decreasing the disease severity with 75% of plant protection. Moreover, ZnO-NPs stimulated the recovery of eggplant as an indicated by improving of morphological and metabolic indicators including plant height(152.5%), root length(106.6%), plant fresh biomass (146%), chlorophyll a (102.8%), chlorophyll b (67.86%), total soluble carbohydrates (48.5%), total soluble protein (81.8%), phenol (10.5%), antioxidant activity and isozymes compared with infected control. Therefore, this study suggests using mycosynthesized ZnO-NPs as an alternative to synthetic fungicides not only to eradicate the Fusarium wilt disease in cultivated eggplant (Solanum melongena) but also to promote the growth parameters and metabolic aspects.

Enhancing tomato plant immune responses to Fusarium wilt disease by red seaweed Jania sp.

The novelty of this study lies in demonstrating a new approach to control wilt diseases using Jania ethyl acetate extract. In the current investigation, the potential impacts of Jania sp. ethyl acetate extract (JE) on Tomato Fusarium oxysporum wilt (FOW) have been studied. The in vitro antifungal potential of JE against F. oxysporum (FO) was examined. GC-MS investigation of the JE revealed that, the compounds possessing fungicidal action were Phenol,2-methoxy-4-(2-propenyl)-,acetate, Eugenol, Caryophyllene oxide, Isoespintanol, Cadinene, Caryophylla-4(12),8(13)-dien-5à-ol and Copaen. Jania sp. ethyl acetate extract exhibited strong antifungal potential against FO, achieving a 20 mmzone of inhibition. In the experiment, two different methods were applied: soil irrigation (SI) and foliar application (FS) of JE. The results showed that both treatments reduced disease index present DIP by 20.83% and 33.33% respectively. The findings indicated that during FOW, proline, phenolics, and the antioxidant enzymes activity increased, while growth and photosynthetic pigments decreased. The morphological features, photosynthetic pigments, total phenol content, and antioxidant enzyme activity of infected plants improved when JE was applied through soil or foliar methods. It is interesting to note that the application of JE had a substantially less negative effect on the isozymes peroxidase and polyphenol oxidase in tomato plants, compared to FOW. These reactions differed depending on whether JE was applied foliarly or via the soil. Finally, the use of Jania sp. could be utilized commercially as an ecologically acceptable method to protect tomato plants against FOW.

Efficient Role of Endophytic Aspergillus terreus in Biocontrol of Rhizoctonia solani Causing Damping-off Disease of Phaseolus vulgaris and Vicia faba.

The wide spread of plant pathogens affects the whole world, threatening national food security. Various fungi including Rhizoctonia solani induce the fungal disease damping-off that negatively affects plant seedlings' growth. Recently, endophytic fungi are used as safe alternatives to chemical pesticides that harm plant and human health. Here, an endophytic Aspergillus terreus was isolated from Phaseolus vulgaris seeds to control damping-off diseases by improving the defense system in Phaseolus vulgaris and Vicia faba seedlings. Endophytic fungus was morphologically and genetically identified Aspergillus terreus, and it is deposited in GeneBank under accession OQ338187. A. terreus demonstrated antifungal efficacy against R. solani with an inhibition zone at 22.0 mm. Moreover, the minimum inhibitory concentrations (MIC) of ethyl acetate extract (EAE) of A. terreus were between 0.3125 and 0.625 mg/mL to inhibit R. solani growth. Precisely 58.34% of the Vicia faba plants survived when A. terreus was added compared with the untreated infected (16.67%). Similarly, Phaseolus vulgaris achieved 41.67% compared to the infected (8.33%). Both groups of treated infected plants showed reduced oxidative damage (reduced Malondialdehyde and hydrogen peroxide levels) as compared to untreated infected plants. Reduced oxidative damage was correlated with the increase in photosynthetic pigments and the antioxidant defense system including polyphenol oxidase, peroxidase, catalase, and superoxide dismutase enzyme activities. Overall, the endophytic A. terreus can be considered an effective tool to control the suppression of Rhizoctonia solani in legumes, especially Phaseolus vulgaris and Vicia faba, as an alternative to synthetic chemical pesticides that harm the environment and human health.

Bio-based antimicrobial food packaging films based on hydroxypropyl starch/polyvinyl alcohol loaded with the biosynthesized zinc oxide nanoparticles.

In the last decades, bio-based active food packaging materials have received much attention. It is known that the utilization of traditional materials for food packaging applications lack some critical characteristics such as resistance to the harmful microbes that cause a damage to the preserved foods. Therefore, the current study aimed to find an alternative packaging films comprises an efficient biopolymers. This research work was designed to prepare film mats using hydroxypropyl starch (HPS), polyvinyl alcohol (PVA), palmitic acid (PA) and biosynthesized zinc oxide nanoparticles (ZnONPs). The fabricated films were coded as 1H, 2H, 3H and 4H based on the utilized concentration of ZnONPs. The biosynthesized ZnONPs and the bio-based films loaded with ZnONPs were characterized. The results revealed that ZnONPs exhibited nearly spherical shape and size ∼40 nm. The surface structure of the produced bioactive packaging films exhibited smooth with homogeneous features, excellent mechanical and thermal stability properties. The prepared bioactive packaging film loaded with ZnONPs (4H) exhibited superior antibacterial activity among other films against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 with inhibition zones 15.1 ± 0.76 and 12.1 ± 0.71 mm respectively. Correspondingly, packing film 4H exhibited potential antifungal activity toward Aspergillus niger RCMB 02724, A. flavus RCMB 02782, Penicillium expansum IMI 89372 and Fusarium oxysporum RCMB 001004 with inhibition zones (16 ± 1.0, 22 ± 0.90, 18.0 ± 1.1 and12.3 ± 0.57 mm respectively). Moreover, all prepared films did not show cytotoxicity on the normal cell line (Wi38) and recorded biodegradability properties that reached around 85 % after four weeks in soil. Based on these results, the antimicrobial films comprising HPS/PVA and loaded with the biosynthesized ZnONPs can be considered as a suitable film for food packaging purposes.

Biostimulation of tomato growth and biocontrol of Fusarium wilt disease using certain endophytic fungi.

BACKGROUND: Tomato plant (Solanum lycopersicum L.) suffers from numerous fungal pathogens that cause damage to yeild production qualitatively and quantitatively. One of the most destructive disease of tomato is Fusarium wilt that caused by soil borne fungus called F. oxysporum. METHODS: In this study, the anti-Fusarium capabilities of the foliar application of fungal endophytes extracts have been investigated on tomato under Fusarium challenges. Antifungal assay, inhibition of conidial germination, disease severity, photosynthetic pigments, osmolytes, secondary metabolites, oxidative stress, peroxidase (POD) and polyphenol oxidases (PPO) isozymes were tested for potential resistance of tomato growing under Fusarium infection. RESULTS: Ethyl acetate extracts of A. flavus MZ045563, A. fumigatus MZ045562 and A. nidulans MZ045561 exhibited antifungal activity toward F. oxysporum where inhibition zone diameters were 15, 12 and 20 mm, respectively. Moreover, extracts of all fungal isolates at concentration 7.5 mg/mL reduced conidia germination from 94.4 to 100%. Fusarium infection caused a destructive effects on tomato plant, high severity desiese index 84.37%, reduction in growth parameters, photosynthetic pigments, and soluble protein. However, contents of proline, total phenol, malondialdehyde (MDA), hydrogen peroxide (H2O2) and antioxidant enzymes activity were increased in tomato plants grown under Fusarium wilt. Treatment of healthy or infected tomato plants by ethyl acetate fungal extracts showed improvements in morphological traits, photosynthetic pigments, osmolytes, total phenol and antioxidant enzymes activity. Besides, the harmful impacts of Fusarium wilt disease on tomato plants have also been reduced by lowering MDA and H2O2 levels. Also, treated tomato plants showed different responses in number and density of POD and PPO isozymes. CONCLUSION: It could be suggested that application of ethyl acetate extracts of tested fungal endophytes especially combination of A. flavus, A. nidulans and A. fumigatus could be commercially used as safe biostimulation of tomato plants as well as biofungicide against tomato Fusarium wilt disease.

Biocontrol of early blight disease of eggplant using endophytic Aspergillus terreus: improving plant immunological, physiological and antifungal activities.

BACKGROUND: The eggplant suffers from many biotic stresses that cause severe damage to crop production. One of the most destructive eggplant pathogens is Alternaria solani, which causes early blight disease. A pot experiment was conducted to evaluate the role of fungal endophytes in protecting eggplant against early blight as well as in improving its growth performance. RESULTS: Endophytic Aspergillus terreus was isolated from Ocimum basilicum leaves and identified morphologically and genetically. In vitro, crude extract of endophytic A. terreus exhibited promising antifungal activity against A. solani where minimum inhibitory concentration (MIC) was 1.25 mg/ml. Severity of the disease and rate of protection from the disease were recorded. Vegetative growth indices, physiological resistance signs (photosynthetic pigments, carbohydrates, proteins, phenols, proline, malondialdehyde (MDA), antioxidant enzymes), and isozymes were estimated. Alternaria solani caused a highly disease severity (87.5%) and a noticeable decreasing in growth characteristics and photosynthetic pigments except for carotenoids. Also, infection with A. solani caused significant decreases in the contents of carbohydrate and protein by 29.94% and 10.52%, respectively. Infection with A. solani caused enhancement in phenolics (77.21%), free proline (30.56%), malondialdehyde (30.26%), superoxide dismutase (SOD) (125.47%), catalase (CAT) (125.93%), peroxidase (POD) (25.07%) and polyphenol oxidase (PPO) (125.37%) compared to healthy plants. In contrast, the use of A. terreus on infected plants succeeded in recovering eggplants from the disease, as the disease severity was recorded (caused protection by 66.67%). Application of A. terreus either on healthy or infected eggplants showed several responses in number and density of peroxidase (POD) and polyphenol oxidase (PPO) isozymes. CONCLUSION: It is necessary for us to address the remarkable improvement in the photosynthetic pigments, protein, carbohydrates, and enzymatic activity compared to infected control, which opens the way for more studies on the use of biocides as safe alternatives against fungal diseases.

Ferric Oxide Colloid: Towards Green Nano-Fertilizer for Tomato Plant with Enhanced Vegetative Growth and Immune Response Against Fusarium Wilt Disease.

Global food crisis due to climate change, pandemic COVID-19 outbreak, and Russia-Ukraine conflict leads to catastrophic consequences; almost 10 percent of the world's population go to bed hungry daily. Narrative solution for green agriculture with high vegetation and crop yield is mandatory; novel nanomaterials can improve plant immunity and restrain plant diseases. Iron is fundamental nutrient element; it plays vital role in enzyme activity and RNA synthesis; furthermore it is involved in photosynthesis electron-transfer chains. This study reports on the facile synthesis of colloidal ferric oxide nanoparticles as novel nano-fertilizer to promote vegetation and to suppress Fusarium wilt disease in tomato plant. Disease index, protection percent, photosynthetic pigments, and metabolic indicators of resistance in plant as response to induction of systemic resistance (SR) were recorded. Results illustrated that Fe2O3 NPs had antifungal activity against F. oxysporum. Fe2O3 NPs (at 20 µg/mL) was the best treatment and reduced percent disease indexes by 15.62 and gave highly protection against disease by 82.15% relative to untreated infected plants. Fe2O3 NPs treatments in either (non-infected or infected) plants showed improvements in photosynthetic pigments, osmolytes, and antioxidant enzymes activity. The beneficial effects of the synthesized Fe2O3 NPs were extended to increase not only photosynthetic pigments, osmolytes contents but also the activities of peroxidase (POD), polyphenol oxidase (PPO), catalase (CAT) and superoxide dismutase (SOD), enzymes of the healthy and infected tomato plants in comparison with control. For, peroxidase and polyphenol oxidase activities it was found that, application of Fe2O3 NPs (10 µg/mL) on challenged plants offered the best treatments which increased the activities of POD by (34.4%) and PPO by (31.24%). On the other hand, application of Fe2O3 NPs (20 µg/mL) on challenged plants offered the best treatments which increased the activities of CAT by (30.9%), and SOD by (31.33%). Supplementary Information: The online version contains supplementary material available at 10.1007/s10904-022-02442-6.

Antifungal Activity of Endophytic Aspergillus terreus Extract Against Some Fungi Causing Mucormycosis: Ultrastructural Study.

Endophytes fungi are applied as favorable safe antifungal agents as well as natural bioactive compounds reservoir. In the current study, the inhibitory effect of endophytic fungus was explained by direct antifungal activity against fungi causing mucormycosis, ultrastructural, and determination of active compounds in fungal extract. Endophytic Aspergillus terreus was isolated from healthy Moringa oleifera leaves and identified morphologically and genetically, and was recorded in gene bank with accession number MW444551.1. Phytochemical analysis and gas chromatography-mass spectroscopy (GC-MS) of ethyl acetate crude extract (EACE) of A. terreus were performed. GC-MS results of EACE of A. terreus revealed that fungal extract contains 16 major bioactive compounds with extensive pharmaceutical activities. Furthermore, EACE of A. terreus revealed a promising antifungal activity against fungi causing mucormycosis as Rhizopus oryzae, Mucor racemosus, and Syncephalastrum racemosum, where inhibition zones of EACE (10 mg/ml) were 20, 37, and 18 mm, respectively. Minimum inhibitory concentration (MIC) of EACE was 0.3125 toward M. racemosus, while 1.25 and 2.5 mg/ml against R. oryzae and S. racemosum, respectively. In the same context, treated R. oryzae, M. racemosus, and S. racemosum with EACE of A. terreus revealed elevation of membrane lipid peroxidation which approves membrane leakage. Furthermore, ultrastructure changes were observed which established alteration in both sporangium and hyphal structures; cell membrane and cytoplasm leakage. In conclusion, endophytic A. terreus has an outstanding antifungal activity against fungi causing mucormycosis.

Antimicrobial, Antioxidant, Cytotoxic Activities and Phytochemical Analysis of Fungal Endophytes Isolated from Ocimum Basilicum.

Fungal endophytes are living inside plants without any harmful effects; the prospecting about them is increased day by day because they can produce bioactive compounds which can be used in different applications. Herein, the current study was aimed to isolate the endophytic fungi from the Ocimum basilicum plant as safe microorganisms and evaluate their biological activities. The results illustrated that three endophytic fungal strains were isolated and identified morphologically and genetically as Aspergillus nidulans, Aspergillus fumigatus, and Aspergillus flavus and deposited in gene bank under accession numbers MZ045561, MZ045562, and MZ045563 respectively. Moreover, cell-free filtrates of endophytic fungal strains were extracted using ethyl acetate, where these crude extracts exhibited promising antimicrobial activity against Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumonia, and Candida albicans at a concentration of 1000 µg/mL. Furthermore, these endophytic strains exhibited a potential antioxidant activity where IC50 of the crude extract of A. nidulans, A. fumigatus, and A. flavus were (166.3, 68.4, and 347.1 µg/mL) and (151.2, 77.9, and 246.3 µg/mL) using DPPH and ABTS methods, respectively. Furthermore, the ethyl acetate crude extracts of these endophytic fungi did not exhibit any cytotoxic effect against Vero and Wi 38 normal cells. GC-MS analysis of the crude extract of A. nidulans, A. fumigatus, and A. flavus indicated the presence of 22, 22, and 20 active compounds, respectively. The major compounds in the fungal extracts are belonging to fatty acids, fatty acid esters, tetrahydrofurans, and sterols. In conclusion, the isolated endophytic A. nidulans, A. fumigatus, and A. flavus from Ocimum basilicum are promising sources for bioactive compounds.

Antagonistic Effect of Plant Growth-Promoting Fungi Against Fusarium Wilt Disease in Tomato: In vitro and In vivo Study.

Fusarium wilt is considered one of the most destructive diseases for tomato plants. The novelty of this work was to investigate the antifungal and plant growth-promoting capabilities of some plant growth-promoting fungi (PGPF). Plant growth-promoting fungi (PGPF) improved the plant health and control plant infections. In this study, two fungal strains as PGPF were isolated and identified as Aspergillus fumigatus and Rhizopus oryzae using molecular method. The extracts of A. fumigatus and R. oryzae exhibited promising antifungal activity against F. oxysporum in vitro. Moreover, antagonistic effect of A. fumigatus and R. oryzae against F. oxysporum causing tomato wilt disease was evaluated in vivo. Disease severity and growth markers were recorded and in vitro antagonistic activity assay of the isolated A. fumigatus and R. oryzae against Fusarium oxysporum was measured. Physiological markers of defense in plant as response to stimulate systemic resistance (SR) were recorded. Our results indicated that A. fumigatus and R. oryzae decreased the percentage of disease severity by 12.5 and 37.5%, respectively. In addition, they exhibited relatively high protection percentage of 86.35 and 59.06% respectively. Fusarium wilt was declined the growth parameters, photosynthetic pigments, total soluble carbohydrate, and total soluble protein, whereas content of free proline, total phenols, and the activity of antioxidant enzymes activity increased under infection. Moreover, application of A. fumigatus and R. oryzae on infected plants successfully recovered the loss of morphological traits, photosynthetic pigment total carbohydrates, and total soluble proteins in comparison to infected control plants. PGPF strains in both non-infected and infected plants showed several responses in number and density of peroxidase (POD) and polyphenol oxidase (PPO) isozymes.

Mycosynthesis of Silica Nanoparticles Using Aspergillus niger: Control of Alternaria solani Causing Early Blight Disease, Induction of Innate Immunity and Reducing of Oxidative Stress in Eggplant.

The threats to the life and production of crops are exacerbated by climate change and the misuse of chemical pesticides. This study was designed to evaluate the effectiveness of biosynthesized silica nanoparticles (SiO2-NPs) as an alternative to pesticides against early blight disease of eggplant. Antifungal activity, disease index, photosynthetic pigments, osmolytes, oxidative stress, antioxidant enzymes activities were tested for potential tolerance of eggplant infected with Alternaria solani. Silica nanoparticles were successfully biosynthesized using Aspergillus niger through green and ecofriendly method. Results revealed that SiO2-NPs exhibited promising antifungal activity against A. solani where MIC was 62.5 µg/mL, and inhibition growth at concentration 1000 µg/mL recorded 87.8%. The disease Index (DI) as a result of infection with A. solani reached 82.5%, and as a result, a severe decrease in stem and root length and number of leaves occurred, which led to a sharp decrease in the photosynthetic pigments. However, contents of free proline, total phenol and antioxidant enzymes activity were increased in infected plants. On the other hand, the treatment with SiO2-NPs 100 ppm led to a great reduction in the disease Index (DI) by 25% and a high protection rate by 69.69%. A clear improvement in growth characteristics and a high content of chlorophyll and total carotenoids was also observed in the plants as a result of treatment with silica nanoparticles in (healthy and infected) plants. Interestingly, the noticeable rise in the content of infected and healthy plants of proline and phenols and an increase in the activity of super oxide dismutase (SOD) and polyphenol oxidase (PPO). It could be suggested that foliar application of SiO2-NPs especially 100 ppm could be commercially used as antifungal and strong inducer of plant physiological immunity against early blight disease.

Daphnia magna and Gammarus pulex, novel promising agents for biomedical and agricultural applications.

Various studies have shown the importance of using different types of Zooplankton biomasses as an additional substance in the diet of fish. In addition, the drainage water of the fish cultures could be used in plant irrigation. In this study, biomasses of water flea Daphnia magna and Gammarus pulex collected and tested, for the first time, their effect against pathogenic microorganisms and on plant germination. The results showed significant antibacterial activity of D. magna and G. pulex against Staphylococcus aureus and Pseudomonas aeruginosa bacteria, as well as antifungal activity against Alternaria solani and Penicillium expansum, which gives the possibility to be used as biocontrol against these bacteria and plant pathogenic fungi. Furthermore, both animals showed positive activity in the germination rate of Vicia faba seed, reaching 83.0 ± 3.5 and 86.0 ± 3.8%, respectively. In conclusion, the biomasses of D. magna and G. pulex are promising and effective agents for their use in the medical field against some pathogenic microbes and as stimulators of plant growth.

Plant Growth-Promoting Fungi as Biocontrol Tool against Fusarium Wilt Disease of Tomato Plant.

Plant growth-promoting fungi (PGPF) improve plant health and resist plant pathogens. The present study was carried out to biocontrol tomato Fusarium wilt using PGPF through antifungal activity and enhance tomato plant immune response. Four PGPF were identified genetically as Aspergillus flavus, Aspergillus niger, Mucor circinelloides and Pencillium oxalicum. In vitro antagonistic activity assay of PGPF against Fusariumoxysporum was evaluated, where it exhibited promising antifungal activity where MIC was in the range 0.25-0.5 mg/mL. Physiological markers of defense in a plant as a response to stimulation of induced systemic resistance (ISR) were recorded. Our results revealed that A. niger, M. circinelloides, A. flavus and P. oxalicum strains significantly reduced percentages of disease severity by 16.60% and 20.83% and 37.50% and 45.83 %, respectively. In addition, they exhibited relatively high protection percentages of 86.35%, 76.87%, 56.87% and 59.06 %, respectively. With concern to the control, it is evident that the percentage of disease severity was about 87.50%. Moreover, the application of M. circinelloides, P. oxalicum, A. niger and A. flavus successfully recovered the damage to morphological traits, photosynthetic pigments' total carbohydrate and total soluble protein of infected plants. Moreover, the application of tested PGPF enhanced the growth of healthy and infected tomato plants.

Inhibition of Aspergillus flavus Growth and Aflatoxin Production in Zea mays L. Using Endophytic Aspergillus fumigatus.

Aspergillus flavus infection of vegetative tissues can affect the development and integrity of the plant and poses dangerous risks on human and animal health. Thus, safe and easily applied approaches are employed to inhibit A. flavus growth. To this end, the fungal endophyte, i.e., Aspergillus fumigatus, was used as a safe biocontrol agent to reduce the growth of A. flavus and its infection in maize seedlings. Interestingly, the safe endophytic A. fumigatus exhibited antifungal activity (e.g., 77% of growth inhibition) against A. flavus. It also reduced the creation of aflatoxins, particularly aflatoxin B1 (AFB1, 90.9%). At plant level, maize seedling growth, leaves and root anatomy and the changes in redox status were estimated. Infected seeds treated with A. fumigatus significantly improved the germination rate by 88.53%. The ultrastructure of the infected leaves showed severe disturbances in the internal structures, such as lack of differentiation in cells, cracking, and lysis in the cell wall and destruction in the nucleus semi-lysis of chloroplasts. Ultrastructure observations indicated that A. fumigatus treatment increased maize (leaf and root) cell wall thickness that consequentially reduced the invasion of the pathogenic A. flavus. It was also interesting that the infected seedlings recovered after being treated with A. fumigatus, as it was observed in growth characteristics and photosynthetic pigments. Moreover, infected maize plants showed increased oxidative stress (lipid peroxidation and H2O2), which was significantly mitigated by A. fumigatus treatment. This mitigation was at least partially explained by inducing the antioxidant defense system, i.e., increased phenols and proline levels (23.3 and 31.17%, respectively) and POD, PPO, SOD and CAT enzymes activity (29.50, 57.58, 32.14 and 29.52%, respectively). Overall, our study suggests that endophytic A. fumigatus treatment could be commercially used for the safe control of aflatoxins production and for inducing biotic stress tolerance of A. flavus-infected maize plants.

Enhancement of Seawater Stress Tolerance in Barley by the Endophytic Fungus Aspergillus ochraceus.

Symbiotic plant-fungi interaction is a promising approach to alleviate salt stress in plants. Moreover, endophytic fungi are well known to promote the growth of various crop plants. Herein, seven fungal endophytes were screened for salt tolerance; the results revealed that Aspergillus ochraceus showed a great potentiality in terms of salt tolerance, up to 200 g L-1. The indole acetic acid (IAA) production antioxidant capacity and antifungal activity of A. ochraceus were evaluated, in vitro, under two levels of seawater stress, 15 and 30% (v/v; seawater/distilled water). The results illustrated that A. ochraceus could produce about 146 and 176 µg mL-1 IAA in 15 and 30% seawater, respectively. The yield of IAA by A. ochraceus at 30% seawater was significantly higher at all tryptophan concentrations, as compared with that at 15% seawater. Moreover, the antioxidant activity of ethyl acetate extract of A. ochraceus (1000 µg mL-1) at 15 and 30% seawater was 95.83 ± 1.25 and 98.33 ± 0.57%, respectively. Crude extracts of A. ochraceus obtained at 15 and 30% seawater exhibited significant antifungal activity against F. oxysporum, compared to distilled water. The irrigation of barley plants with seawater (15 and 30%) caused notable declines in most morphological indices, pigments, sugars, proteins, and yield characteristics, while increasing the contents of proline, malondialdehyde, and hydrogen peroxide and the activities of antioxidant enzymes. On the other hand, the application of A. ochraceus mitigated the harmful effects of seawater on the growth and physiology of barley plants. Therefore, this study suggests that the endophytic fungus A. ochraceus MT089958 could be applied as a strategy for mitigating the stress imposed by seawater irrigation in barley plants and, therefore, improving crop growth and productivity.

Bacillus megaterium-Mediated Synthesis of Selenium Nanoparticles and Their Antifungal Activity against Rhizoctonia solani in Faba Bean Plants.

Rhizoctonia root-rot disease causes severe economic losses in a wide range of crops, including Vicia faba worldwide. Currently, biosynthesized nanoparticles have become super-growth promoters as well as antifungal agents. In this study, biosynthesized selenium nanoparticles (Se-NPs) have been examined as growth promoters as well as antifungal agents against Rhizoctonia solani RCMB 031001 in vitro and in vivo. Se-NPs were synthesized biologically by Bacillus megaterium ATCC 55000 and characterized by using UV-Vis spectroscopy, XRD, dynamic light scattering (DLS), and transmission electron microscopy (TEM) imaging. TEM and DLS images showed that Se-NPs are mono-dispersed spheres with a mean diameter of 41.2 nm. Se-NPs improved healthy Vicia faba cv. Giza 716 seed germination, morphological, metabolic indicators, and yield. Furthermore, Se-NPs exhibited influential antifungal activity against R. solani in vitro as well as in vivo. Results revealed that minimum inhibition and minimum fungicidal concentrations of Se-NPs were 0.0625 and 1 mM, respectively. Moreover, Se-NPs were able to decrease the pre-and post-emergence of R. solani damping-off and minimize the severity of root rot disease. The most effective treatment method is found when soaking and spraying were used with each other followed by spraying and then soaking individually. Likewise, Se-NPs improve morphological and metabolic indicators and yield significantly compared with infected control. In conclusion, biosynthesized Se-NPs by B. megaterium ATCC 55000 are a promising and effective agent against R. solani damping-off and root rot diseases in Vicia faba as well as plant growth inducer.