Unveiling Fungi Armor: Preliminary Study on Fortifying Pisum sativum L. Seeds against Drought with Schizophyllum commune Fries 1815 Polysaccharide Fractions.

Amidst worsening climate change, drought stress imperils global agriculture, jeopardizing crop yields and food security, thereby necessitating the urgent exploration of sustainable methods like biopriming for the harnessing of beneficial microorganisms to bolster plant resilience. Recent research has revealed diverse biological compounds with versatile applications produced by Schizophyllum commune, rendering this fungus as a promising contender for biopriming applications. For the first time, this study aimed to investigate the potential of S. commune exo- (EPSH) and intra-polysaccharides (IPSH) isolated from two strains-Italian (ITA) and Serbian (SRB)-under submerged cultivation to enhance the resilience of Pisum sativum L. seeds through the biopriming technique. Testing of the seed quality for the bioprimed, hydroprimed, and unprimed seeds was conducted using a germination test, under optimal and drought conditions, while characterization of the PSHs included FTIR analysis, microanalysis, and determination of total protein content (TPC). The FTIR spectra of EPSH and IPSH were very similar but revealed the impurities, while microanalysis and TPC confirmed a different presence of proteins in the isolated PSHs. In optimal conditions, the IPSH SRB increased germination energy by 5.50% compared to the control; however, the highest percentage of germination (94.70%) was shown after biopriming with the PSH isolated from the ITA strain. Additionally, all assessed treatments resulted in a boost in seedling growth and biomass accumulation, where the ITA strain demonstrated greater effectiveness in optimal conditions, while the SRB strain showed superiority in drought conditions. The drought tolerance indices increased significantly in response to all examined treatments during the drought, with EPSH ITA (23.00%) and EPSH SRB (24.00%) demonstrating the greatest effects. Results of this preliminary study demonstrate the positive effect of isolated PSH, indicating their potential as biopriming agents and offering insights into novel strategies for agricultural resilience.

Bioactivity Profiling of Daedaleopsis confragosa (Bolton) J. Schröt. 1888: Implications for Its Possible Application in Enhancing Women's Reproductive Health.

This study investigates the bioactivity profile of wood-rotting fungal species Daedaleopsis confragosa (Bolton) J. Schröt. 1888, focusing on its antioxidant, cytotoxic, and genotoxic activities and enzyme modulation properties with respect to its possible application in terms of enhancing women's reproductive health. Two types of extracts, including those based on EtOH extraction (DC) and hydrodistillation (DCHD), were investigated. The results indicate that the radical scavenging capacity against the DPPH radical and reduction potential were stronger in the DC extracts owing to the higher total phenolic content (TPC) and total flavonoid content (TFC) (25.30 ± 1.05 mg GAE/g d.w. and 2.84 ± 0.85 mg QE/g d.w., respectively). The same trend was observed in the protein phosphatase-1 (PP1) activity and in the genotoxic activity against the δ virus since only the DC extract exhibited DNA disintegration regarding a dilution of 1:100. Conversely, the DCHD extract exhibited increased hemolytic and cytotoxic effects (339.39% and IC50 = 27.76 ± 0.89 µg/mL-72 h incubation, respectively), along with greater inhibition of the AChE enzyme (IC50 = 3.11 ± 0.45 mg/mL) and hemolytic activity. These results suggest that terpenoids and steroids may be responsible for the observed activity in DCHD as these compounds could potentially be extracted following the HD procedure. This comprehensive bioactivity profiling offers valuable insights into the potential therapeutic applications of D. confragosa from Serbia and underscores the importance of further investigations for harnessing its pharmacological potential.

Antioxidant proficiency in Serbian mushrooms: a comparative study on Hydnum repandum L. 1753 from mycorrhizal and edible niches.

This study aimed to evaluate the antioxidant potential of autochthonous Hydnum repandum through LC-MS/MS profiling, total phenolic content (TP), total protein content (TPR), and antioxidant capabilities (DPPH, ABTS, and FRAP assays) across various extracts (CHCl3, acetone, 70% EtOH, 80% MeOH, and hot water). LC-MS/MS analysis revealed a predominant presence of quinic acid in polar solvents (ranging from 531.37 to 676.07 ng/mL), while EtOH and MeOH extracts exhibited elevated total phenolic levels (27.44 ± 0.32 and 28.29 ± 3.62 mg GAE/g d.w., respectively). Impressively, H. repandum showcased remarkable antioxidant properties, as evidenced by its FRAP values (57.29 to 199.96 mg AAE/g d.w.), ABTS values (5.69 to 29.95 mg TE/g d.w.), and IC50 values in the DPPH assay (91.40 to 372.55 µg/mL), which exhibited a strong correlation with TP. Notably, the acetone extract exhibited the most robust antioxidant activity where the highest TPR was observed, suggesting synergism of primary and secondary metabolites.

Do Ganoderma Species Represent Novel Sources of Phenolic Based Antimicrobial Agents?

Ganoderma species have been recognized as potential antimicrobial (AM) agents and have been used in traditional Chinese medicine (TCM) for a long time. The aim of this study is to examine the AM potential of autochthonous Ganoderma species (G. applanatum, G. lucidum, G. pfeifferi and G. resinaceum) from Serbia. The extraction of fungal material was prepared in different solvents (ethanol-EtOH, water-H2O, chloroform-CHCl3). Antibacterial activity (ABA) was determined using disk-diffusion, agar-well diffusion, and micro-dilution method, while for antifungal properties disk-diffusion and pour plate method were applied. Antiviral activity was tested on model DNA virus LK3 and determined by plaque assay. Statistical PCA analysis was applied for detection of correlation effects of phenolics and AM activities, while LC-MS/MS was performed for phenolics quantification. G. resinaceum CHCl3 extract expressed the most potent ABA against P. aeruginosa (MIC = 6.25 mg/mL), probably due to presence of flavonoids and 2,5-dihydroxybenzoic acid. Among H2O extracts, the highest ABA was determined for G. pfeifferi against both E. coli and S. aureus (21 and 19 mm, respectively). EtOH extracts of G. pfeifferi and G. resinaceum were the most effective against A. niger (23.8 and 20.15 mm, respectively), with special impact of phenolic acids and flavonoid isorhamnetin, while C. albicans showed the lowest susceptibility. The most potent antiviral inhibitor was G. lucidum (70.73% growth inhibition) due to the high amount of phenolic acids. To the best of our knowledge, this is the first report of a methodical AM profile of G. pfeifferi and G. resinaceum from the Balkan region including PCA analysis.

Effects of UV Stress in Promoting Antioxidant Activities in Fungal Species Тrametes versicolor (L.) Lloyd and Flammulina velutipes (Curtis) Singer.

Trametes versicolor and Flammulina velutipes, after submerged cultivation, with intermittent exposure to ultraviolet light (UV), were studied for Total Phenolic content (TP) and Total Proteins (TPR) contents and antioxidant properties against free radicals. The TP and TPR were determined by a spectrophotometric method and Lowry's assay, respectively. Liquid chromatography with mass spectrometry (LC-MS/MS) was used to quantify polyphenols. Different in vitro assays determined the antioxidant activities of the extracts. Mycelia extract from F. velutipes after 14 days and filtrate from T. versicolor after 21 days of incubation gave the highest TP 59.60 ± 0.14 and 50.03 ± 0.66 mg GAE/g d.w., respectively. Mycelia extract from T. versicolor after 28 days of incubation had the highest TPR (183.53 ± 2.84 mg BSAE/g d.w.). The LC-MS/MS analysis indicated that p-hydroxybenzoic and protocatechuic acids are the most abundant. Trametes versicolor filtrate after 14 days and F. velutipes filtrate after 21 days (71.29 ± 0.54% and 73.5 ± 1.81, respectively) had the highest scavenging activity in SOA. Correlation analysis indicated that all extracts' antioxidant (AO) potential strongly correlated with TP (R2 = 0.83-1.0). The data confirmed that stress factors such as UV exposure could stimulate the production of secondary metabolites and natural AOs, especially phenolic acids in test fungi.

Polyamines in Edible and Medicinal Fungi from Serbia: A Novel Perspective on Neuroprotective Properties.

The therapeutic effectiveness of current neurodegenerative disease treatments is still under debate because of problems with bioavailability and a range of side effects. Fungi, which are increasingly recognized as sources of natural antioxidants and acetylcholinesterase (AChE) enzyme inhibitors, may thus serve as potent neuroprotective agents. Previous studies have associated the anti-AChE and antioxidant activities of fungi mostly with polysaccharides and phenolic compounds, while other secondary metabolites such as polyamines (PAs) have been neglected. This study aimed to investigate eight edible and medicinal fungi from Serbia, marking the initial investigation into the neuroprotective capabilities of Postia caesia, Clitocybe odora, Clitopilus prunulus, and Morchella elata. Neuroprotective activity was examined using the Ellman assay, while the antioxidant capacity was tested by conducting DPPH, NO, ABTS, and FRAP tests. PA levels were determined by high-performance liquid chromatography (HPLC) coupled with fluorescent detection. Ganoderma applanatum and Lepista nuda exhibited the most robust anti-AChE (98.05 ± 0.83% and 99.94 ± 3.10%, respectively) and antioxidant activities, attributed to the synergistic effects of the total protein, total phenolic, and PA levels. Furthermore, P. caesia displayed significant AChE inhibition (88.21 ± 4.76%), primarily linked to the elevated spermidine (SPD) (62.98 ± 3.19 mg/kg d.w.) and putrescine (PUT) levels (55.87 ± 3.16 mg/kg d.w.). Our results highlight the need for thorough research to comprehend the intricate relationships between distinct fungus species and AChE inhibition. However, it is important to recognize that more research is required to identify the precise substances causing the reported inhibitory effects.

Antimicrobial and Toxic Effects of Boswellia serrata Roxb. and Mentha piperita Linn. Essential Oils on Vaginal Inhabitants.

Commercial essential oils (EOs) of incense, Boswellia serrata Roxb, and mint, Mentha piperita L., were investigated against vaginal bacterial and Candida albicans isolates for antimicrobial potential and safety use. The antimicrobial activity of EOs was investigated through a double-dilution micro-plate assay. A brine shrimp assay was used for the determination of toxicity, while the determination of the chemical composition of EOs was carried out using GS-MS. Obtained minimal inhibitory (MIC) and minimal bactericidal concentration (MBC) point to the activity of mint essential oil (EO) against the multi-resistant P. aeruginosa isolate (MIC/MBC at 6.25 µL/mL), while MIC and MBC values for other isolates were reached at higher concentrations (25-50 µL/mL). According to the toxicity assay, the incense EO reached the LC50 value at 3.07 µL/mL, while mint EO showed higher toxicity at lower concentrations (0.5 µL/mL) and the LC50 could not be determined. The highest antimicrobial potential was obtained for incense against P. aeruginosa. Although the toxicity assay showed high toxicity of mint EO to the eggs of aquatic crustaceans Artemia salina, further testing of EO toxicity is proposed, for example on healthy cell-lines. According to the GC/MS spectrometry, the most represented components of mint EO were the oxygenated hydrocarbons L-menthone (20.86%) and menthol (31.86%), and they could be proposed for further antimicrobial and toxicity investigation.

Mushroom Species Stereum hirsutum as Natural Source of Phenolics and Fatty Acids as Antioxidants and Acetylcholinesterase Inhibitors.

Many lignicolous mushroom species are used as a food supplement and may represent an alternative treatment of Alzheimer's disease (AD). This study aimed to evaluate acetylcholinesterase inhibition (AChEI) of Stereum hirsutum together with antioxidant activity (AO) and cytotoxic activity against HepG2 cells. Different extracts (water, ethanol, methanol, polysaccharide) were analyzed, with respect to their mineral composition and chemical content. Ethanol extract was the most potent in AChEI (98.44 %) and demonstrated cytotoxic activity (91.96 % at 900.00 µg/mL), while the highest AO was demonstrated for polar extracts (methanol and water) as well. These activities may be attributed to determined phenolics (hydroxybenzoic and quinic acid) and fatty acids (FA), while biflavonoid amentoflavone may be responsible for cytotoxic activity. The most prevalent FA was linoleic (40.00 %) and the domination of unsaturated FA (UFA) (71.91 %) over saturated (26.96 %) was observed. This is the first report of AChEI of S. hirsutum extracts and first detection of amentoflavone. Due to high amount of UFA and well-expressed AChEI, this species can be considered as a potent food supplement in the palliative therapy of AD.

Comparison of Two Schizophyllum commune Strains in Production of Acetylcholinesterase Inhibitors and Antioxidants from Submerged Cultivation.

In recent years, fungi have been recognized as producers of acetylcholinesterase (AChE) inhibitors, agents important for the prevention of Alzheimer's disease (AD). This study aimed to examine the AChE inhibitory, the antioxidative and antibacterial activity of two different Schizophyllum commune strains that originated from Serbia (SRB) and Italy (IT). Submerged cultivation of grown mycelia (M) and fermentation broth (F) of ethanol (EtOH) and polysaccharide (PSH) extracts lasted for 7, 14, 21 and 28 days. For AChE activity Ellman method was performed, while for antioxidative activity, sevendifferent assays were conducted: DPPH, ABTS, FRAP, SOA, OH, NO together with total phenolic content. Antimicrobial screen, LC-MS/MS technique and FTIR measurements were performed. Different isolates exhibited different AChE activity, with PSH being the strongest (SRB, M, 28 days IC90 79.73 ± 26.34 µg/mL), while in EtOH extracts, IT stood out (F, 14 days, IC50 0.8 ± 0.6 µg/mL). PSH extracts (7 days) exhibit significant antioxidative activity (AO), opposite to EtOH extracts where 14 and 21days periods stood out. Only tw extracts showed antibacterial activity. Following LC-MS/MS analysis p-hydroxybenzoic and gallic acids were the most abundant phenolics. PSH extracts demonstrated remarkable results, making this study debut and introducing S. commune as a valuable resource of AChE inhibitors.