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.

Polycyclic aromatic hydrocarbon (PAH) phytoaccumulation in urban areas by Platanus × acerifolia, Celtis australis, and Tilia grandifolia leaves and branches.

Polycyclic aromatic hydrocarbon (PAH) concentrations in the leaves and 1-year-old branches of three common tree species growing in a middle-sized city located in a moderate climate zone were estimated. For this purpose, PAH phytoaccumulation in Platanus × acerifolia, Celtis australis, and Tilia grandifolia species from highly urbanized, traffic congested, and highly PAH-contaminated streets was compared with trees from non-contaminated parks in the same urban core. The gathered data was used to define 17 PAH profiles, identify the main PAH pollution emission sources, and determine the organ and species specificity of PAHs accumulation. Due to the direct absorption of polluted air via stomata, the leaves accumulated up to 30% more PAHs compared to the 1-year-old branches. As expected, PAH concentrations were much higher in street trees, while heavy weight PAHs (with five and six rings) were accumulated in the highest concentrations. The highest foliar Σ17 PAH concentrations were detected in street-grown C. australis, followed by P. acerifolia and T. grandifolia (502.68, 488.45, and 339.47 ng g-1 dry weight (DW), respectively). The same pattern was noted for Σ17 PAHs in branches (414.89, 327.58, and 342.99 ng g-1 DW, respectively). Thus, T. grandifolia emerged as the least effective PAH sink as it accumulated up to ~ 40% less PAHs than P. acerifolia and C. australis leaves/branches. Among the 17 tracked PAHs, benzo[a]anthracene, benzo[a]pyrene, dibenzo[a,h]anthracene, and pyrene were found to have accumulated in the highest concentrations in all analyzed species irrespective of the site, and accounted for more than 50% of the total detected PAHs. Finally, a "black box" about species and organ specificity, as well as specific drivers that limit PAHs uptake capacity by trees, was opened, while this work provides insights into further PAH phytoremediation strategies.

Variations in Proline Content, Polyamine Profiles, and Antioxidant Capacities among Different Provenances of European Beech (Fagus sylvatica L.).

International provenance trials are a hot topic in forestry, and in light of climate change, the search for more resilient beech provenances and their assisted migration is one of the challenges of climate-smart forestry. The main aim of the study was to determine intraspecific variability in European beech (Fagus sylvatica L.) among 11 beech provenances according to total antioxidant capacities estimated by various assays, such as DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid), FRAP (ferric reducing antioxidant power) assay, and radical scavenging capacity against nitric oxide (RSC-NO assays), as well as osmolyte content, primarily individual polyamines (putrescine, spermidine, and spermine), and free proline content. Polyamine amounts were quantified by using HPLC coupled with fluorescent detection after dansylation pretreatment. The highest values for radical scavenger capacity assays (ABTS, DPPH, and FRAP) were measured in the German provenances DE47 and DE49. Also, the highest NO inhibition capacity was found in the provenance DE49, while the highest content of proline (PRO), total phenolic content (TPC), and total flavonoid content (TFC) was recorded in DE47. The Austrian AT56 and German provenance DE49 were most abundant in total polyamines. This research underlines the importance of the application of common antioxidant assays as well as osmolyte quantification as a criterion for the selection of climate-ready beech provenances for sustainable forest management.

In vitro selection of drought-tolerant white poplar clones based on antioxidant activities and osmoprotectant content.

Introduction: In light of upcoming climate change, there is an urgent requirement for tree improvement regarding adaptability to drought-caused stress and the development of quick and reliable screening methodologies for genotypes' drought tolerance. White poplar is, despite its high adaptability, considered to be an endangered tree species in Serbia, which gives it special importance in the preservation and improvement of biodiversity of riparian ecosystems. The main goal of this research was to evaluate the tolerance of five white poplar clones to the presence of polyethylene glycol (PEG 6000 molecular weight 6000) (different concentrations (e.g. 0 g/L, 1 g/L, 10 g/L, 20 g/L, and 50 g/L) in Aspen Culture Medium (ACM). Methods: The tolerance of the clones was evaluated by using morphological parameters (shoot fresh and dry weight, root fresh and dry weight), photosynthetic pigments (contents of chlorophyll a, chlorophyll b, carotenoids, and chlorophyll a+b), and biochemical parameters (total phenolic content, total flavonoid content, ferric reducing antioxidant power, antioxidant activities (DPPH activity and ABTS assay), free proline content and glycine betaine content. Results and Discussion: The values of morphological and photosynthetic pigments declined with an increase in the concentration of PEG 6000. At a concentration of 50 g/L, the content of shoot fresh mass decreased by 41%, the content of Chl a by 68%, Chl b by 65%, and Car by 76% compared to the control. Also, at the same medium, there was an increase in the content of total phenols, accumulation of proline, the content of glycine betaine as well as in antioxidant activity. Based on the obtained results, it can be assumed that more drought-tolerant clones are characterized by high values for biomass, high content of photosynthetic pigments, and high content of proline and glycine betaine in conditions similar to drought in vitro. Clone L-80 showed better results in most of the tested parameters, especially compared to the reference clone Villafranca.

A Fine-Tuning of the Plant Hormones, Polyamines and Osmolytes by Ectomycorrhizal Fungi Enhances Drought Tolerance in Pedunculate Oak.

The drought sensitivity of the pedunculate oak (Quercus robur L.) poses a threat to its survival in light of climate change. Mycorrhizal fungi, which orchestrate biogeochemical cycles and particularly have an impact on the plant's defense mechanisms and metabolism of carbon, nitrogen, and phosphorus, are among the microbes that play a significant role in the mitigation of the effects of climate change on trees. The study's main objectives were to determine whether ectomycorrhizal (ECM) fungi alleviate the effects of drought stress in pedunculate oak and to investigate their priming properties. The effects of two levels of drought (mild and severe, corresponding to 60% and 30% of field capacity, respectively) on the biochemical response of pedunculate oak were examined in the presence and absence of ectomycorrhizal fungi. To examine whether the ectomycorrhizal fungi modulate the drought tolerance of pedunculate oak, levels of plant hormones and polyamines were quantified using UPLC-TQS and HPLC-FD techniques in addition to gas exchange measurements and the main osmolyte amounts (glycine betaine-GB and proline-PRO) which were determined spectrophotometrically. Droughts increased the accumulation of osmolytes, such as proline and glycine betaine, as well as higher polyamines (spermidine and spermine) levels and decreased putrescine levels in both, mycorrhized and non-mycorrhized oak seedlings. In addition to amplifying the response of oak to severe drought in terms of inducible proline and abscisic acid (ABA) levels, inoculation with ECM fungi significantly increased the constitutive levels of glycine betaine, spermine, and spermidine regardless of drought stress. This study found that compared to non-mycorrhized oak seedlings, unstressed ECM-inoculated oak seedlings had higher levels of salicylic (SA) and abscisic acid (ABA) but not jasmonic acid (JA), indicating a priming mechanism of ECM is conveyed via these plant hormones. According to a PCA analysis, the effect of drought was linked to the variability of parameters along the PC1 axe, such as osmolytes PRO, GB, polyamines, and plant hormones such as JA, JA-Ile, SAG, and SGE, whereas mycorrhization was more closely associated with the parameters gathered around the PC2 axe (SA, ODPA, ABA, and E). These findings highlight the beneficial function of the ectomycorrhizal fungi, in particular Scleroderma citrinum, in reducing the effects of drought stress in pedunculate oak.

Spermidine dietary supplementation and polyamines level in reference to survival and lifespan of honey bees.

Honey bee health has been an important and ongoing topic in recent years. Honey bee is also an important model organism for aging studies. Polyamines, putrescine, spermidine and spermine, are ubiquitous polycations, involved in a wide range of cellular processes such as cell growth, gene regulation, immunity, and regulation of lifespan. Spermidine, named longevity elixir, has been most analysed in the context of aging. One of the several proposed mechanisms behind spermidine actions is antioxidative activity. In present study we showed that dietary spermidine supplementation: (a) improved survival, (b) increased the average lifespan, (c) influenced the content of endogenous polyamines by increasing the level of putrescine and spermidine and decreasing the level of spermine, (d) reduced oxidative stress (MDA level), (e) increased the antioxidant capacity of the organism (FRAP), (f) increased relative gene expression of five genes involved in polyamine metabolism, and (g) upregulated vitellogenin gene in honey bees. To our knowledge, this is the first study on honey bee polyamine levels in reference to their longevity. These results provide important information on possible strategies for improving honey bee health by introducing spermidine into their diet. Here, we offer spermidine concentrations that could be considered for that purpose.

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.

Metal- and Organ-Specific Response to Heavy Metal-Induced Stress Mediated by Antioxidant Enzymes' Activities, Polyamines, and Plant Hormones Levels in Populus deltoides.

Besides anthropogenic factors, climate change causes altered precipitation patterns that indirectly affect the increase of heavy metals in soils due to hydrological effects and enhanced leaching (i.e., Cd and Ni), especially in the vicinity of mines and smelters. Phytoextraction is a well-known, powerful "green" technique for environmental clean-up that uses plants to extract, sequester, and/or detoxify heavy metals, and it makes significant contributions to the removal of persistent inorganic pollutants from soils. Poplar species, due to their growth features, high transpiration rate, large biomass, and feasible reproduction represent great candidates for phytoextraction technology. However, the consequences of concomitant oxidative stress upon plant metabolism and the mechanism of the poplar's tolerance to heavy metal-induced stress are still not completely understood. In this study, cuttings of poplar species (Populus deltoides W. Bartram ex Marshall) were separately exposed to two heavy metals (Cd2+ and Ni2+) that were triple the maximum allowed amount (MAA) (according to national legislation). The aim of the study was to estimate the effects of heavy metals on: (I) the accumulation of free and conjugated polyamines, (II) plant hormones (including abscisic acid-ABA and indole-3-acetic acid-IAA), and (III) the activities of different antioxidant enzymes at root and leaf levels. By using the selected ion monitoring (SIM) mode of gas chromatography with mass spectrometry (GC/MS) coupled with the isotopically labeled technique, amounts of ABA and IAA were quantified, while polyamine amounts were determined by using high-performance liquid chromatography (HPLC) with fluorometric detection after derivatization. The results showed that P. deltoides responded to elevated concentrations of heavy metals in soils by exhibiting metal- and organ-specific tolerance. Knowledge about tolerance mechanisms is of great importance for the development of phytoremediation technology and afforestation programs for polluted soils.

Ectomycorrhizal Fungi Modulate Pedunculate Oak's Heat Stress Responses through the Alternation of Polyamines, Phenolics, and Osmotica Content.

The physiological and biochemical responses of pedunculate oaks (Quercus robur L.) to heat stress (HS) and mycorrhization (individually as well in combination) were estimated. One-year-old Q. robur seedlings were grown under controlled conditions in a pot experiment, inoculated with a commercial inoculum of ectomycorrhizal (ECM) fungi, and subjected to 72 h of heat stress (40 °C/30 °C day/night temperature, relative humidity 80%, photoperiod 16/8 h) in a climate chamber, and they were compared with seedlings that were grown at room temperature (RT). An in-depth analysis of certain well-known stress-related metrics such as proline, total phenolics, FRAP, ABTS, non-protein thiols, and lipid peroxidation revealed that mycorrhized oak seedlings were more resistant to heat stress (HS) than non-mycorrhized oaks. Additionally, levels of specific polyamines, total phenolics, flavonoids, and condensed tannins as well as osmotica (proline and glycine betaine) content were measured and compared between four treatments: plants inoculated with ectomycorrhizal fungi exposed to heat stress (ECM-HS) and those grown only at RT (ECM-RT) versus non-mycorrhized controls exposed to heat stress (NM-HS) and those grown only at room temperature (NM-RT). In ectomycorrhiza inoculated oak seedlings, heat stress led to not only a rise in proline, total phenols, FRAP, ABTS, non-protein thiols, and lipid peroxidation but a notable decrease in glycine betaine and flavonoids. Amounts of three main polyamines (putrescine, spermine, and spermidine) were quantified by using high-performance liquid chromatography coupled with fluorescent detection (HPLC/FLD) after derivatization with dansyl-chloride. Heat stress significantly increased putrescine levels in non-mycorrhized oak seedlings but had no effect on spermidine or spermine levels, whereas heat stress significantly increased all inspected polyamine levels in oak seedlings inoculated with ectomycorrhizal inoculum. Spermidine (SPD) and spermine (SPM) contents were significantly higher in ECM-inoculated plants during heat stress (approximately 940 and 630 nmol g-1 DW, respectively), whereas these compounds were present in smaller amounts in non-mycorrhized oak seedlings (between 510 and 550 nmol g-1 DW for Spd and between 350 and 450 nmol g-1 DW for Spm). These findings supported the priming and biofertilizer roles of ectomycorrhizal fungi in the mitigation of heat stress in pedunculate oaks by modification of polyamines, phenolics, and osmotica content.

Species-Level Differences in Osmoprotectants and Antioxidants Contribute to Stress Tolerance of Quercus robur L., and Q. cerris L. Seedlings under Water Deficit and High Temperatures.

The general aim of this work was to compare the leaf-level responses of different protective components to water deficit and high temperatures in Quercus cerris L. and Quercus robur L. Several biochemical components of the osmotic adjustment and antioxidant system were investigated together with changes in hormones. Q. cerris and Q. robur seedlings responded to water deficit and high temperatures by: (1) activating a different pattern of osmoregulation and antioxidant mechanisms depending on the species and on the nature of the stress; (2) upregulating the synthesis of a newly-explored osmoprotectant, dimethylsulphoniopropionate (DMSP); (3) trading-off between metabolites; and (4) modulating hormone levels. Under water deficit, Q. cerris had a higher antioxidant capacity compared to Q. robur, which showed a lower investment in the antioxidant system. In both species, exposure to high temperatures induced a strong osmoregulation capacity that appeared largely conferred by DMSP in Q. cerris and by glycine betaine in Q. robur. Collectively, the more stress-responsive compounds in each species were those present at a significant basal level in non-stress conditions. Our results were discussed in terms of pre-adaptation and stress-induced metabolic patterns as related to species-specific stress tolerance features.

Developing Forest Therapy Programmes Based on the Health Benefits of Terpenes in Dominant Tree Species in Tara National Park (Serbia).

Numerous medical studies have shown the positive effects of forests on different aspects of human health. This study deals with the content of major terpenes in dominant coniferous species in Tara National Park, Serbia, in order to explore the potential for the development of a novel health tourism programme based on forest therapy. Main terpenes were analysed using a headspace-sampling technique coupled with gas-chromatography-mass spectrometry (Head-space-GC/MS). Needles of fir and spruce growing in the vicinity of hiking trails were investigated for possibilities to perform such therapy. Major detected terpenes were α-cadinol and spathulenol previously described as antiviral, antitumor, antimicrobial and immunomodulatory agents. The results of the study were favourable and worked well with the existing walking infrastructure in the observed area of the Tara Mountain, as they act as invaluable resources for designing the structured forest bathing walks. The study not only adds to the knowledge in the environmental and public health realm but also to tourism and sustainability studies.

Citric acid as soil amendment in cadmium removal by Salix viminalis L., alterations on biometric attributes and photosynthesis.

During the past decade, the target in cleaning polluted sites is an application of chelating agents, such as citric acid (CA), which is proposed as a good candidate in the promotion of phytoremediation. Among heavy metals, cadmium (Cd) is one of the most common and dangerous elements, which strongly disturbs morphophysiological properties in plants. A pot experiment was assessed to evaluate the influence of CA in Cd phytoremediation in alkaline soil by Salix viminalis (clone SV068). The effects of CA on Cd bioavailability, mobility, and distribution in plants, various morphometric measurements, and physiological performances as photosynthesis, transpiration, water use efficiency, and pigment content were tested. The highest Cd accumulation was evident after 60 days of growing, in plants subjected to combined treatment of CA with a higher dose of Cd. Application of CA showed a beneficial effect in maintaining the photosynthetic rate as well as gas exchange capacity in willows grown in Cd-contaminated soil. Furthermore, CA slightly increased plant growth and biomass production, depending on applied Cd dose and harvest period. A chelating agent like CA showed strong influence in plant response to combat Cd toxicity.

Radionuclides in some edible and medicinal macrofungal species from Tara Mountain, Serbia.

Edible and medicinal macrofungi used in human diet represent not only important sources of nutritive elements but toxic substances as well (heavy metals and radionuclides). Radioactivity levels of four radionuclides ((40)K, (137)Cs, (226)Ra, (228)Ra) were determined in the basidiomata (fruiting bodies of a Basidiomycetes) of six lignicolous (Fomitopsis pinicola, Ganoderma applanatum, Hericium clathroides, Megacollybia platyphylla, Pluteus cervinus, Trametes gibbosa) and three mycorrhizal (Boletus luridus, Boletus sp. 1, Boletus sp. 2) species as well as their soil (wood) substrates by gamma spectrometry (high-resolution high-purity germanium (HPGe) detector). The aim was to investigate their ability for radionuclide absorption according to transfer factors (from soil and wood), to predict potential bioindicator species as well as species with potential risk for human use. Samples were taken during years 2011 and 2012, at two sites in forest ecosystem of Tara Mountain (Serbia). Observed concentration ranges per dry weight were as follows: 29-3,020 Bq/kg ((40)K), 21.9-735 Bq/kg ((137)Cs), 3-39 Bq/kg ((226)Ra), and 2.0-18 Bq/kg ((228)Ra). Obtained results indicate that the type of basidiome (fleshy/tough), most likely due to a different metabolic rate, has a very important role in radionuclide accumulation. The highest activity concentrations of all analyzed radionuclides were found in species with fleshy basidiomata--P. cervinus, H. clathroides, M. platyphylla, and Boletus species. A species-specific influence on radionuclide uptake was more prominent comparing to habitat differences and the role of fungal trophic mode. No significant variations were observed regarding radionuclide activity among the same fungal species from different sampling sites.