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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 (TiO 2 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 TiO 2 NPs by reducing TiCl 4 salt into TiO 2 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 TiO 2 NPs, and the diameter of the inhibitory zone increased with increasing TiO 2 NPs concentration. In an in vivo assay, barley plants treated with T. cf. asperellum  or TiO 2 NPs were used to evaluate their biocontrol efficiency against  B. sorokiniana,  in which  T. cf. asperellum  and TiO 2 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 TiO 2 NPs in response to B. sorokiniana treatment were quantitively estimated. Hence,  T. cf. asperellum and TiO 2 NPs improve the plant’s tolerance and reduce the growth inhibitory effect of B. sorokiniana . Conclusion Subsequently, T. cf. asperellum and TiO 2 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 TiO 2 NPs to the management of spot blotch disease in barley plants, and ultimately to the enhancement of barley plant quality and productivity. Key points • T. cf. asperellum showed strong antagonistic activity against  B. sorokiniana invitro . • Plant- based synthesis of TiO 2 NPs (10- 25 nm) using Aloe vera L. aqueous leaf extract. • B. sorokiniana triggers morphological and biochemical changes in barley plants, causing spot blotch disease. • T. cf. asperellum or green synthesized TiO 2 NPs positively increased the host plant's tolerance against this disease by inducing of osmolytes and antioxidant defense-related enzyme production.

Mycosynthesis of nanoparticle (NP) production is a potential ecofriendly technology for large scale production. In the present study, copper oxide nanoparticles (CuONPs) have been synthesized from the live cell filtrate of the fungus Penicillium chrysogenum. The created CuONPs were characterized via several techniques, namely Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), transmission electron microscope (TEM), scanning electron microscope (SEM), and energy-dispersive X-ray spectroscopy (EDX). Furthermore, the biosynthesized CuONPs were performed against biofilm forming Klebsiella oxytoca ATCC 51,983, Escherichia coli ATCC 35,218, Staphylococcus aureus ATCC 25,923, and Bacillus cereus ATCC 11,778. The anti-bacterial activity result was shown with the zone of inhibition determined to be 14 ± 0.31 mm, 16 ± 0.53 mm, 11 ± 0.57 mm, and 10 ± 0.57 mm respectively. Klebsiella oxytoca and Escherichia coli were more susceptible to CuONPs with minimal inhibitory concentration (MIC) values 6.25 and 3.12 µg/mL, respectively, while for Staphylococcus aureus and Bacillus cereus, MIC value was 12.5 and 25 μg/mL, respectively. The minimum biofilm inhibition concentration (MBIC) result was more evident, that the CuONPs have excellent anti-biofilm activity at sub-MIC levels reducing biofilm formation by 49% and 59% against Klebsiella oxytoca and Escherichia coli, while the results indicated that the MBIC of CuONPs on Bacillus cereus and Staphylococcus aureus was higher than 200 μg/mL and 256 μg/mL, respectively, suggesting that these CuONPs could not inhibit mature formatted biofilm of Bacillus cereus and Staphylococcus aureus in vitro. Overall, all the results were clearly confirmed that the CuONPs have excellent anti-biofilm ability against Klebsiella oxytoca and Escherichia coli. The prepared CuONPs offer a smart approach for biomedical therapy of resistant microorganisms because of its promoted antimicrobial action, but only for specified purposes.

Mycobacterium bovis Bacillus Calmette–Guérin (BCG) has long been used as a vaccine against tuberculosis and bladder cancer. BCG is also an effective adjuvant; it has long been known that its cell wall acts as an adjuvant. Recent research has revealed that various lipids and glycolipids are involved in the induction of innate immunity as ligands for C-type lectin receptors. However, the effects of the components of BCG used as adjuvants are not comparable to those of live BCG. On the other hand, since live bacteria are essential for the protective effect of BCG, a hypothesis has been proposed that proteins actively secreted by living bacteria are responsible for important immune responses, and various secreted proteins have been characterized. However, this hypothesis has not yet been verified. In this review, we discuss the purification, identification, and characterization of BCG-secreted proteins, namely culture filtrate proteins (CFP), and mention new possibilities for CFP.

Paclitaxel is the top-selling anticancer medicine in the world. In vitro culture of Corylus avellana has been made known as a promising and inexpensive strategy for producing paclitaxel. Fungal elicitors have been named as the most efficient strategy for enhancing the biosynthesis of secondary metabolites in plant cell culture. In this study, endophytic fungal strain HEF17 was isolated from C. avellana and identified as Camarosporomyces flavigenus . C. avellana cell suspension culture (CSC) elicited with cell extract (CE) and culture filtrate (CF) derived from strain HEF17, either individually or combined treatment, in mid and late log phase was processed for modeling and optimizing growth and paclitaxel biosynthesis regarding CE and CF concentration levels, elicitor adding day, and CSC harvesting time using multilayer perceptron-genetic algorithm (MLP-GA). The results displayed higher accuracy of MLP-GA models (0.89–0.95) than regression models (0.56–0.85). The great accordance between the predicted and observed values of output variables (dry weight, intracellular, extracellular and total yield of paclitaxel, and also extracellular paclitaxel portion) for both training and testing subsets supported the excellent performance of developed MLP-GA models. MLP-GA method presented a promising tool for selecting the optimal conditions for maximum paclitaxel biosynthesis. An Excel® estimator, HCC-paclitaxel, was designed based on MLP-GA model as an easy-to-use tool for predicting paclitaxel biosynthesis in C. avellana CSC responding to fungal elicitors.

• Receptor-like cytoplasmic kinase subfamily VII (RLCK-VII) proteins are the central immune kinases in plant pattern-recognition receptor (PRR) complexes, and they orchestrate a complex array of defense responses against bacterial and fungal pathogens. However, the role of RLCK-VII in plant–oomycete pathogen interactions has not been established. Phytophthora capsici is a notorious oomycete pathogen that infects many agriculturally important vegetables. • Here, we report the identification of RXLR25, an RXLR effector that is required for the virulence of P. capsici. In planta expression of RXLR25 significantly enhanced plants’ susceptibility to Phytophthora pathogens. Microbial pattern-induced immune activation in Arabidopsis was severely impaired by RXLR25. We further showed that RXLR25 interacts with RLCK-VII proteins. • Using nine rlck-vii high-order mutants, we observed that RLCK-VII-6 and RLCK-VII-8 members are required for resistance to P. capsici. The RLCK-VII-6 members are specifically required for Phytophthora culture filtrate (CF)-induced immune responses. RXLR25 directly targets RLCK-VII proteins such as BIK1, PBL8, and PBL17 and inhibits pattern-induced phosphorylation of RLCK-VIIs to suppress downstream immune responses. • This study identified a key virulence factor for P. capsici, and the results revealed the importance of RLCK-VII proteins in plant–oomycete interactions.

Background Salinity is a major threat to the agriculture industry due to the negative impact of salinity stress on crop productivity. In the present study, we isolated rhizobacteria and evaluated their capacities to promote crop growth under salt stress conditions. Results We isolated rhizospheric bacteria from sand dune flora of Pohang beach, Korea, and screened them for plant growth-promoting (PGP) traits. Among 55 bacterial isolates, 14 produced indole-3-acetic acid (IAA), 10 produced siderophores, and 12 produced extracellular polymeric and phosphate solubilization. Based on these PGP traits, we selected 11 isolates to assess for salinity tolerance. Among them, ALT29 and ALT43 showed the highest tolerance to salinity stress. Next, we tested the culture filtrate of isolates ALT29 and ALT43 for IAA and organic acids to confirm the presence of these PGP products. To investigate the effects of ALT29 and ALT43 on salt tolerance in soybean, we grew seedlings in 0 mM, 80 mM, 160 mM, and 240 mM NaCl treatments, inoculating half with the bacterial isolates. Inoculation with ALT29 and ALT43 significantly increased shoot length (13%), root length (21%), shoot fresh and dry weight (44 and 35%), root fresh and dry weight (9%), chlorophyll content (16–24%), Chl a (8–43%), Chl b (13–46%), and carotenoid (14–39%) content of soybean grown under salt stress. Inoculation with ALT29 and ALT43 also significantly decreased endogenous ABA levels (0.77-fold) and increased endogenous SA contents (6–16%), increased total protein (10–20%) and glutathione contents, and reduced lipid peroxidation (0.8–5-fold), superoxide anion (21–68%), peroxidase (12.14–17.97%), and polyphenol oxidase (11.76–27.06%) contents in soybean under salinity stress. In addition, soybean treated with ALT29 and ALT43 exhibited higher K + uptake (9.34–67.03%) and reduced Na + content (2–4.5-fold). Genes involved in salt tolerance, GmFLD19 and GmNARK , were upregulated under NaCl stress; however, significant decreases in GmFLD19 (3–12-fold) and GmNARK (1.8–3.7-fold) expression were observed in bacterial inoculated plants. Conclusion In conclusion, bacterial isolates ALT29 and ALT43 can mitigate salinity stress and increase plant growth, providing an eco-friendly approach for addressing saline conditions in agricultural production systems.

The demand for high-quality strawberries continues to grow, emphasizing the need for innovative agricultural practices to enhance both yield and fruit quality. In this context, the utilization of natural products, such as biostimulants, has emerged as a promising avenue for improving strawberry production while aligning with sustainable and eco-friendly agricultural approaches. This study explores the influence of a bacterial filtrate (BF), a vegetal-derived protein hydrolysate (PH), and a standard synthetic auxin (SA) on strawberry, investigating their effects on yield, fruit quality, mineral composition and metabolomics of leaves and fruits. Agronomic trial revealed that SA and BF significantly enhanced early fruit yield due to their positive influence on flowering and fruit set, while PH treatment favored a gradual and prolonged fruit set, associated with an increased shoot biomass and sustained production. Fruit quality analysis showed that PH-treated fruits exhibited an increase of firmness and soluble solids content, whereas SA-treated fruits displayed lower firmness and soluble solids content. The ionomic analysis of leaves and fruits indicated that all treatments provided sufficient nutrients, with heavy metals within regulatory limits. Metabolomics indicated that PH stimulated primary metabolites, while SA and BF directly affected flavonoid and anthocyanin biosynthesis, and PH increased fruit quality through enhanced production of beneficial metabolites. This research offers valuable insights for optimizing strawberry production and fruit quality by harnessing the potential of natural biostimulants as viable alternative to synthetic compounds.

Fusarium species are considered one of the most destructing plant pathogens. In the current study, bimetallic zinc oxide-copper oxide nanoparticles (ZnO-CuO NPs) were myco-synthesized using Aspergillus fumigatus for controlling Fusarium oxysporum growth. Aspergillus fumigatus was isolated from soil and identified morphologically and genetically. The myco-synthesized ZnO-CuO NPs were characterized using UV-Vis, DLS, HR-TEM, SEM, and XRD analyses. HR-TEM characterization method indicated that, the biosynthesized bimetallic ZnO-CuO NPs appeared as semi-spherical with the average diameter specified as 54.18 ± 1.9 nm. The DLS method described the characteristic particle size diffusion and was calculated as 85.52 nm, 90.85 nm, and 92.85 nm for ZnO NPs, CuO NPs, and ZnO-CuO NPs, respectively. Additionally, the SEM image of ZnO-CuO NPs displays basic NP surface character and the exterior impression was apparent. The biosynthesized ZnO-CuO NPs were separated naturally as spherical particles connected within the fungal filtrate, which displays as illuminated NPs fused and capped with the fungal filtrate. Antifungal activity of bimetallic ZnO-CuO NPs was evaluated against F. oxysporum . Results revealed that bimetallic ZnO-CuO NPs exhibited promising antifungal activity toward F. oxysporum where inhibition zone at 1000 µg/ml was 22.8 ± 0.76 mm, and MIC was 125 µg/ml. Moreover, growth inhibition percentages of F. oxysporum at different concentrations of bimetallic ZnO-CuO NPs 1000, 500, 250, and 125 µg/ml were 88.9, 65.5, 41.1, and 8.9% respectively, where the highest inhibition was 88.9% at concentration 1000 µg/ml, while the lowest inhibition was 8.9% at concentration 125 µg/ml. In TEM ultrastructure results, the treated F. oxysporum with ZnO-CuO NPs, a clear destruction was found in all cell contents and disintegration of the cell wall as well as destruction of the plasma membrane. Also, the nucleus appeared as small size and damaged shape and the chromatin materials distributed with several dark stained bodies in cytoplasm. In conclusion, bimetallic ZnO-CuO NPs were successfully myco-synthesized using A. fumigatus , where it had promising antifungal activity against F. oxysporum .

The arecanut palm is one of the most important industrial crops in tropical area around the world. The root rot of arecanut palm, which is caused by Cerrena unicolor, has led to heavy economic losses and restricted greatly the development of arecanut industry, especially in Hainan province of China. The common use of chemical agents has worsened the problems of the emergence of resistant pathogens and the pollution of agricultural environment. This study aims to screen and identify a more effective and environment friendly biocontrol method for the prevention and treatment of root rot of arecanut palm. The mycelium growth rate is investigated to select antagonistic bacteria from tropical crop rotation fields which show improved resistance against soil-borne pathogens, and the strain P42 is revealed with the strongest antagonistic effects (82.18%). Based on 16 s rDNA sequence analysis, the strain P42 is identified as Lysinibacillus boronitolerans. In vitro antimicrobial activity shows that the strain P42 exhibits broad-spectrum antagonistic activity against a wide variety of tropical agricultural fungal pathogens, including Cerrena unicolor, Magnaporthe oryzea, Botryodiplodia theobromae, Neoscytalidium dimidiatum, Thanatephorus cucumeris, Fusarium oxysporum, and Botrytis cinerea Per.. The antagonistic activity of the culture of P42 is tolerant to common proteases, longer storage time, and temperature range of 40–121 °C; and is significantly influenced by alkaline (7–9) and acidic (1–2) pH, as well as by ultraviolet ray treatment for more than 30 min. The investigation on the antagonistic activity of the crude extract of fermentation filtrate indicates that the active compounds might be lipopeptides, polyketones, or proteins. To our knowledge, this is the first report of L. boronitolerans as potential bio-reagents for controlling root rot of arecanut palm caused by Cerrena unicolor.

Addressing the pressing issues of increased food demand, declining crop productivity under varying agroclimatic conditions, and the deteriorating soil health resulting from the overuse of agricultural chemicals, requires innovative and effective strategies for the present era. Microbial bioformulation technology is a revolutionary, and eco-friendly alternative to agrochemicals that paves the way for sustainable agriculture. This technology harnesses the power of potential microbial strains and their cell-free filtrate possessing specific properties, such as phosphorus, potassium, and zinc solubilization, nitrogen fixation, siderophore production, and pathogen protection. The application of microbial bioformulations offers several remarkable advantages, including its sustainable nature, plant probiotic properties, and long-term viability, positioning it as a promising technology for the future of agriculture. To maintain the survival and viability of microbial strains, diverse carrier materials are employed to provide essential nourishment and support. Various carrier materials with their unique pros and cons are available, and choosing the most appropriate one is a key consideration, as it substantially extends the shelf life of microbial cells and maintains the overall quality of the bioinoculants. An exemplary modern bioformulation technology involves immobilizing microbial cells and utilizing cell-free filters to preserve the efficacy of bioinoculants, showcasing cutting-edge progress in this field. Moreover, the effective delivery of bioformulations in agricultural fields is another critical aspect to improve their overall efficiency. Proper and suitable application of microbial formulations is essential to boost soil fertility, preserve the soil’s microbial ecology, enhance soil nutrition, and support crop physiological and biochemical processes, leading to increased yields in a sustainable manner while reducing reliance on expensive and toxic agrochemicals. This manuscript centers on exploring microbial bioformulations and their carrier materials, providing insights into the selection criteria, the development process of bioformulations, precautions, and best practices for various agricultural lands. The potential of bioformulations in promoting plant growth and defense against pathogens and diseases, while addressing biosafety concerns, is also a focal point of this study.