Enhancing drought tolerance in Malva parviflora plants through metabolic and genetic modulation using Beauveria bassiana inoculation.

BACKGROUND: Enhancing crops' drought resilience is necessary to maintain productivity levels. Plants interact synergistically with microorganisms like Beauveria bassiana to improve drought tolerance. Therefore, the current study investigates the effects of biopriming with B. bassiana on drought tolerance in Malva parviflora plants grown under regular irrigation (90% water holding capacity (WHC)), mild (60% WHC), and severe drought stress (30% WHC). RESULTS: The results showed that drought stress reduced the growth and physiological attributes of M. parviflora. However, those bioprimed with B. bassiana showed higher drought tolerance and enhanced growth, physiological, and biochemical parameters: drought stress enriched malondialdehyde and H2O2 contents. Conversely, exposure to B. bassiana reduced stress markers and significantly increased proline and ascorbic acid content under severe drought stress; it enhanced gibberellic acid and reduced ethylene. Bioprimed M. parviflora, under drought conditions, improved antioxidant enzymatic activity and the plant's nutritional status. Besides, ten Inter-Simple Sequence Repeat primers detected a 25% genetic variation between treatments. Genomic DNA template stability (GTS) decreased slightly and was more noticeable in response to drought stress; however, for drought-stressed plants, biopriming with B. bassiana retained the GTS. CONCLUSION: Under drought conditions, biopriming with B. bassiana enhanced Malva's growth and nutritional value. This could attenuate photosynthetic alterations, up-regulate secondary metabolites, activate the antioxidant system, and maintain genome integrity.

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.

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.

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.

Prospective of mycorrhiza and Beauvaria bassiana silica nanoparticles on Gossypium hirsutum L. plants as biocontrol agent against cotton leafworm, Spodoptera littoralis.

BACKGROUND: Plant-herbivorous insects are a severe danger to the world's agricultural production of various crops. Insecticides used indiscriminately resulted in habitat destruction due to their high toxicity, as well as disease resistance. In this respect, the development of a sustainable approach to supreme crop production with the least damage is a crucially prerequisite. As a result, the current study was carried out to understand the potential effect of arbuscular mycorrhizal (AM) fungi along with Beauvaria bassiana silica nanoparticles (Si NPs) as a new approach to increase cotton (Gossypium hirsutum L. Merr.) defense against an insect herbivore, Spodoptera littoralis. AM and non-AM cotton plants were infested with S. littoralis and then sprayed with a biopesticide [B. bassiana Si NPs] or a chemical insecticide (Chlorpyrifos). RESULTS: The gas chromatography-mass spectrometry (GC-MS) analysis of B. bassiana Si NPs fungal extract showed that the major constituents identified were Oleyl alcohol, trifluoroacetate, 11-Dodecen-1-AL and 13-Octadecenal, (Z)-(CAS). Besides, results revealed a highly significant decrease in growth parameters in S. littoralis infested plants, however, with AM fungal inoculation a substantial improvement in growth traits and biochemical parameters such as protein and carbohydrates contents was observed. In addition, stimulation in proline and antioxidant enzymes activity and a decrease in malondialdehyde content were observed after AM inoculation. CONCLUSION: AM fungi mitigate the harmful effects of herbivorous insects by strengthening the cotton plant's health via enhancing both morphological and biochemical traits that can partially or completely replace the application of chemical insecticides.

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.

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.

Arbuscular mycorrhizal fungi and Trichoderma viride cooperative effect on biochemical, mineral content, and protein pattern of onion plants.

The benefits of growth-stimulating microbes in crop production represent great opportunities for recent agricultural practices. Thus, the present investigation deals with examining whether arbuscular mycorrhizal (AM) fungi or Trichoderma viride application or their dual inoculation could improve the biochemical parameters and mineral and nutrient contents of onion plants (Allium cepa) under glasshouse conditions. The results evidenced that both AM fungi and T. viride are compatible with each other, and their combined use is effective, not only in improving the biochemical parameters, such as total soluble carbohydrates, protein contents, total free amino acids, acid, and alkaline phosphatases, but also in increasing mineral and nutrient contents (N, P, K+ , Ca2+ , Mg2+ , and Zn) in onion plants, in which an increase of 67%, 49%, and 112% was observed in shoot onion P content with AM, T. viride, and with their dual inoculation, respectively, as compared with the controlled ones. Also, AM fungal colonization percentage augmented greatly with T. viride inoculation. Moreover, the protein profile of onion leaves revealed the appearance of newly protein bands with AM and T. viride applications. Therefore, their applications improved onion plant development, which could be used to replace the expensive chemical fertilizers, thus increasing onion quality.

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.

Alleviation of the adverse effects of NaCl stress on tomato seedlings (Solanum lycopersicum L.) by Trichoderma viride through the antioxidative defense system.

BACKGROUND: Trichoderma viride are well known for their biocontrol capabilities, but little is known about how they stimulate plant development and increase their resistance to salt stress. One of the main abiotic factors limiting crop development and yield is salt stress. Therefore, the purpose of this work was to ascertain how NaCl effects on T. viride growth as well as on the seedlings morphological and physio-biochemical parameters of tomato (Solanum lycopersicum L.) under plate culture conditions. Additionally, a pot experiment was conducted to determine how T. viride affected the development characteristics of tomato plants subjected to various salt concentrations (50 and 100 mM NaCl). T. viride's contribution to tomato seedling stress tolerance was also closely examined. RESULTS: Results showed that 100 mM NaCl decreased the colony diameter of T. viride by 13.4% compared to the control. Under plate and greenhouse conditions, tomato seedlings exposed to salt exposure exhibited an overall decline in growth. Also, a reduction in relative water content (RWC) and protein contents occurred under salt stress. At the same time, increases were found in proline, total phenolics, flavonoids, H2O2 content, malondialdehyde, likewise the activities of peroxidase (POD), catalase (CAT), polyphenol oxidase (PPO), and ascorbate peroxidase (APX) enzymes. Even though, with T. viride application, the salt negative effects on both morphological and physio-biochemical parameters were mitigated to a greater extent. T. viride increased proline and total antioxidant capacity (TAC) in tomato seedlings at 100 mM NaCl by an average of 20.66 and 43.82% compared to their comparable control. T. viride increased the activities of CAT, PPO, and APX enzymes by 74.6, 58.48, and 61.61% at 50 mM NaCl compared to non-saline control seedlings. As well, T. viride decreased MDA and H2O2 contents by an average of 14 and 24.8% in tomato seedlings at 50 mM NaCl compared to their comparable control. Also, under 100 mM NaCl, the T. viride-treated tomato seedlings showed increased total phenolics (17.85%) and flavonoids (33.17%) compared to non- treated one. CONCLUSION: Hence, our research sheds new insight on the pathways by which T. viride can boost tomato seedling tolerance to salt stress at morphological and physio-biochemical levels by activating both enzymatic and non-enzymatic antioxidant defense systems.

Evaluation of the Antifungal Activity of Bacillusamyloliquefaciens and B. velezensis and Characterization of the Bioactive Secondary Metabolites Produced against Plant Pathogenic Fungi.

Endophytic bacteria are plant-beneficial bacteria with a broad host range. They provide numerous benefits to their hosts, helping them tolerate several biotic and abiotic stresses. An interest has recently been developed in endophytic bacteria which are producing bioactive compounds that contribute to the biological control of various phytopathogens. This research paper aimed to investigate the potentiality of new local strains of endophytic bacteria such as Bacillus amyloliquefaciens and B. velezensis and the production of several antimicrobial metabolites associated with the biocontrol of Alternaria sp., which cause serious diseases and affect important vegetable crops in Egypt. Twenty-five endophytic bacteria isolates were obtained from different plants cultivated in El-Sharkia Governorate, Egypt. Dual culture technique was used to evaluate the bacterial isolates' antagonistic potentiality against Alternaria sp. and Helminthosporium sp. The most active bacterial isolates obtained were selected for further screening. The antifungal activity of the most active endophytic bacterial isolate was assessed in vivo on pepper seedlings as a biocontrol agent against Alternaria sp. A significant antifungal activity was recorded with isolates C1 and T5 against Alternaria sp. and Helminthosporium sp. The bacterial endophyte discs of C1 and T5 showed the highest inhibitory effect against Alternaria sp. at 4.7 and 3.1 cm, respectively, and Helminthosporium sp. at 3.9 and 4.0 cm, respectively. The most active endophytic isolates C1 and T5 were identified and the 16S rRNA sequence was submitted to the NCBI GenBank database with accession numbers: MZ945930 and MZ945929 for Bacillus amyloliquefaciens and Bacillus velezensis, respectively. The deformity of pathogenic fungal mycelia of Alternaria sp. and Helminthosporium sp. was studied under the biotic stress of bacteria. The culture filtrates of B. amyloliquefaciens and B. velezensis were extracted with different solvents, and the results indicated that hexane was the most efficient. Gas Chromatography-Mass Spectrometry revealed that Bis (2-ethylhexyl) phthalate, Bis (2-ethylhexyl) ester, and N,N-Dimethyldodecylamine were major constituents of the endophytic crude extracts obtained from B. amyloliquefaciens and B. velezensis. The in vivo results showed that Alternaria sp. infection caused the highest disease incidence, leading to a high reduction in plant height and in the fresh and dry weights of pepper plants. With B. amyloliquefaciens application, DI significantly diminished compared to Alternaria sp. infected pepper plants, resulting in an increase in their morphological parameters. Our findings allow for a reduction of chemical pesticide use and the control of some important plant diseases.

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.