The regulatory roles of a plant neurotransmitter, acetylcholine, on growth, PSII photochemistry and antioxidant systems in wheat exposed to cadmium and/or mercury stress.

Heavy metals increase in nature due to anthropogenic activities and negatively impact the growth, progress, and efficiency of plants. Among the toxic metal pollutants that can cause dangerous effects when accumulated by plants, mercury (Hg) and cadmium (Cd) were investigated in this study. These metals typically inhibit important enzymes and halt their functioning, thereby adversely affecting the capability of plants to achieve photosynthesis, respiration, and produce quality crops. Acetylcholine (ACh) serves as a potent neurotransmitter present in both primitive and advanced plant species. Its significant involvement in diverse metabolic processes, particularly in regulating growth and adaptation to stress, needs to be further elucidated. For this aim, effects of acetylcholine (ACh1, 10 µM; ACh2, 100 µM) were survey in Triticum aestivum under Hg and/or Cd stress (Hg, 50 µM; Cd, 100 µM). Wheat seedlings exhibited a growth retardation of about 24% under Hg or Cd stress. Combined stress conditions (Cd + Hg) resulted in a decrease in RWC by approximately 16%. Two different doses of ACh treatment to stressed plants positively affected growth parameters and regulated the water relations. Gas exchange was limited in stress groups, and the photochemical quantum competency of PSII (Fv/Fm) was suppressed. Cd + ACh1 and Cd + ACh2 treatments resulted in approximately 2-fold and 1.5-fold improvement in stomatal conductance and carbon assimilation rate, respectively. Similarly, improvement was observed with ACh treatments in wheat seedlings under Hg stress. Under Cd and/or Hg stress, high levels of H2O2 accumulated and lipid peroxidation occurred. According to our results, ACh treatment upon Cd and Hg stresses improved the activities of SOD, POX, and APX, thereby reducing oxidative damage. In conclusion, ACh treatment was found to ensure stress tolerance and limit the adverse effects caused by heavy metals.

Metabolomic Profiles and Biopharmaceutical Properties of Petrosimonia brachiata and P. nigdeensis from Turkey.

Halophytic plants possess a huge range of active constituents and medicinal benefits. In this study, extracts (water, ethanol, ethyl acetate, dichloromethane, and n-hexane) of two halophytes of the genus Petrosimonia (P. brachiata and P. nigdeensis) were investigated for their phytochemical profiles and pharmacological properties. The phytochemical profiles of both species were investigated using an untargeted metabolomics approach based on high-resolution mass spectrometry. The two species show different polyphenolic profiles and these are influenced by the different extraction solvents used. The same extracts were used for different bioactivity assays. The results show that all extracts yielded total flavonoid and phenolic contents of 11.14-24.22 mg GAE/g and 3.15-22.03 mg RE/g, respectively. While extracts of both species demonstrated a radical scavenging ability in the ABTS assay (16.12-98.02 mg TE/g), only the polar and moderately polar extracts (water, ethanol, and ethyl acetate) showed scavenging potential in the DPPH assay (4.74-16.55 mg TE/g). A reducing potential was also displayed by all extracts in the CUPRAC and FRAP assays (26.02-80.35 mg TE/g and 31.70-67.69 mg TE/g, respectively). The total antioxidant capacity of the extracts ranged from 0.24 to 2.17 mmol TE/g, and the metal chelating activity ranged from 14.74 to 33.80 mg EDTAE/g. The water extracts possessed a higher metal chelating power than the other extracts. All extracts acted as inhibitors of acetylcholinesterase (0.16-3.85 mg GALAE/g) and amylase (0.11-1.28 mmol ACAE/g). Moreover, apart from the water extracts, the other extracts also showed anti-butyrylcholinesterase activity (0.73-2.86 mg GALAE/g), as well as anti-tyrosinase (36.74-61.40 mg KAE/g) and anti-glucosidase (2.37-2.73 mmol ACAE/g) potential. In general, the water extracts were found to be weak inhibitors of the tested enzymes, while the ethanol extracts mostly showed an inhibitory effect. The obtained findings revealed the antioxidant and enzyme inhibitory properties of these two species and demonstrated that the solvent type used affected the pharmacological properties of the extracts and hence, can be useful to further investigate the active constituents yielded in the extracts and understand the mechanisms involved.

UHPLC-ESI-QTOF-MS metabolite profiles of different extracts from Pelargonium endlicherianum parts and their biological properties based on network pharmacological approaches.

In the present study, we aimed to investigate the chemical profiles and biological activities of different extracts (ethyl acetate, dichloromethane, ethanol, and water) of Pelargonium endlicherianum parts (aerial parts and roots). Free radical scavenging, reducing power, phosphomolybdenum, and metal chelating were assayed for antioxidant properties. To detect enzyme inhibitory properties, cholinesterase, amylase, glucosidase, and tyrosinase were chosen as target enzymes. The ethanol extract of the aerial parts contained higher amounts of total bioactive compounds (120.53 mg GAE/g-24.46 mg RE/g). The ethanol and water extracts of these parts were tentatively characterized by UHPLC-ESI-QTOF-MS and 95 compounds were annotated. In addition, the highest acetylcholiesterase (3.74 mg GALAE/g) and butyrylcholinesterase (3.92 mg GALAE/g) abilities were observed by the ethanol extract of roots. The water extract from aerial parts exhibited the most pronounced inhibitory effects on multiple cancer cell lines, especially A549 (IC50: 23.2 µg/mL) and HT-29 (IC50: 27.43 µg/mL) cells. Using network pharmacology, P. endlicherianum compounds were studied against cancer, revealing well-connected targets such as epidermal growth factor receptor (EGFR), phosphoinositide-3-kinase (PI3K), AKT, receptor tyrosine-protein kinase erbB-2, and growth factor receptor bound protein 2 (GRB2) with significant impact on cancer-related pathways. The results could open a new path from natural treasure to functional applications with P. endlicherianum and highlight a new study on other uninvestigated Pelargonium species.

Hesperidin and chlorogenic acid mitigate arsenic-induced oxidative stress via redox regulation, photosystems-related gene expression, and antioxidant efficiency in the chloroplasts of Zea mays.

The ubiquitous metalloid arsenic (As), which is not essential, can be found extensively in the soil and subterranean water of numerous nations, raising substantial apprehensions due to its impact on both agricultural productivity and sustainability. Plants exposed to As often display morphological, physiological, and growth-related abnormalities, collectively leading to reduced productivity. Polyphenols, operating as secondary messengers within the intricate signaling networks of plants, assume integral functions in the acquisition of resistance to diverse environmental stressors, including but not limited to drought, salinity, and exposure to heavy metals. The pivotal roles played by polyphenols in these adaptive processes underscore their profound significance in plant biology. This study aims to elucidate the impact of hesperidin (HP) and chlorogenic acid (CA), recognized as potent bioactive compounds, on maize plants exposed to As. To achieve this objective, the study examined the physiological and biochemical impacts, including growth parameters, photosynthesis, and chloroplastic antioxidants, of HP (100 µM) and CA (50 µM) on Zea mays plants exposed to arsenate stress (AsV, 100 µM - Na2HAsO4⋅7H2O). As toxicity led to reductions in fresh weight (FW) and dry weight (DW) by 33% and 26%, respectively. However, the application of As+HP and As + CA increased FW by 22% and 40% and DW by 14% and 17%, respectively, alleviating the effects of As stress. As toxicity resulted in the up-regulation of PSII genes (psbA and psbD) and PSI genes (psaA and psaB), indicating a potential response to the re-formation of degraded regions, likely driven by the heightened demand for photosynthesis. Exogenous HP or/and CA treatments effectively counteracted the adverse effects of As toxicity on the photochemical quantum efficiency of PSII (Fv/Fm). H2O2 content showed a 23% increase under As stress, and this increase was evident in guard cells when examining confocal microscopy images. In the presence of As toxicity, the chloroplastic antioxidant capacity can exhibit varying trends, with either a decrease or increase observed. After the application of CA and/or HP, a significant increase was observed in the activity of GR, APX, GST, and GPX enzymes, resulting in decreased levels of H2O2 and MDA. Additionally, the enhanced functions of MDHAR and DHAR have modulated the redox status of ascorbic acid (AsA) and glutathione (GSH). The HP or CA-mediated elevated levels of AsA and GSH content further contributed to the preservation of redox homeostasis in chloroplasts facing stress induced by As. In summary, the inclusion of HP and CA in the growth medium sustained plant performance in the presence of As toxicity by regulating physiological and biochemical characteristics, chloroplastic antioxidant enzymes, the AsA-GSH cycle and photosynthesis processes, thereby demonstrating their significant potential to confer resistance to maize through the mitigation of As-induced oxidative damage and the safeguarding of photosynthetic mechanisms.

Extractions of aerial parts of Hippomarathrum scabrum with conventional and green methodologies: Chemical profiling, antioxidant, enzyme inhibition, and anti-cancer effects.

Hippomarathrum scabrum L. is an endemic medicinal plant in Turkey; however, there have been few studies investigating the phytochemistry and biological properties of these plants has not been investigated. The aim of this work is to determine the chemical composition of different extracts (extracts obtained by using supercritical carbon dioxide extraction, accelerated solvent extraction, homogenizer-assisted extraction, microwave-assisted extraction, and ultrasound-assisted extraction from Hippomarathrum scabrum L., and evaluate their biological properties. The analysis revealed that 5-O-caffeoylquinic acid, rutin, and isorhamnetin 3-O-rutinoside were the main bioactive compounds. The extract obtained by accelerated extraction contains the highest concentration of 5-O-Caffeoylquinic acid (7616.74 ± 63.09 mg/kg dry extract) followed by the extract obtained by homogenizer-assisted extraction (6682.53 ± 13.04 mg/kg dry extract). In antioxidant tests, all extracts expressed significant antioxidant activity. Also, cytotoxic and anticancer effects of these plant extracts were detected in the human prostate cancer cell line. Intrinsic apoptotic genes were up-regulated and anti-apoptotic genes were down-regulated in human prostate cancer cells after inhibition concentration dose treatment. The findings are promising, and suggest the use of these plant extracts could be used as natural sources with different biological activities, as well as anticancer agents.

Polyvinylpyrrolidone-coated copper nanoparticles dose-dependently conferred tolerance to wheat under salinity and/or drought stress by improving photochemical activity and antioxidant system.

Copper (Cu) is one of the essential micronutrients for plants and has been used extensively in agricultural applications from the past to the present. However, excess copper causes toxic effects such as inhibiting photosynthesis, and disrupting biochemical processes in plants. Nanotechnology applications have offered a critical method for minimizing adverse effects and improving the effectiveness of copper nanoparticles. For this purpose, this study investigated the physiological and biochemical effects of polyvinylpyrrolidone (PVP)-coated Cu nanoparticles (PVP-Cu NP, N1, 100 mg L-1; N2, 400 mg L-1) in Triticum aestivum under alone or combined with salt (S, 150 mM NaCl) and/or drought (D, %10 PEG-6000) stress. Salinity and water deprivation caused 51% and 22% growth retardation in wheat seedlings. The combined stress condition (S + D) resulted in an approximately 3-fold reduction in the osmotic potential of the leaves. PVP-Cu NP treatments to plants under stress, especially N1 dose, were effective in restoring growth rate and regulating water relations. All stress treatments limited gas exchange in stomata and suppressed the maximal quantum yield of PSII (Fv/Fm). More than 50% improvement was observed in stomatal permeability and carbon assimilation rate under S + N1 and S + N2 applications. Examination of OJIP transient parameters revealed that N1 treatments protected photochemical reactions by reducing the dissipated energy flux (DIo/RC) in drought and S + D conditions. Exposure to S and/or D stress caused high hydrogen peroxide (H2O2) accumulation and lipid peroxidation in wheat leaves. The results indicated that S + N1 and S + N2 treatments reduced oxidative damage by stimulating the activities of antioxidant enzymes superoxide dismutase (SOD), peroxidase (POX), and ascorbate peroxidase (APX). Although similar effects were observed at D and S + D conditions with 100 mg L-1 PVP-Cu NP treatments (N1), the curative effect of the N2 dose was not observed. In D + N1 and S + D + N1 groups, AsA regeneration and GSH redox status were maintained by triggering APX, GR, and other enzyme activities belonging to the AsA-GSH cycle. In these groups, N2 treatment did not contribute to the availability of enzymatic and non-enzymatic antioxidants. As a result, this study revealed that N1 dose PVP-Cu NP application was successful in providing stress tolerance and limiting copper-induced adverse effects under all stress conditions.

Graphene oxide-based aerogel stimulates growth, mercury accumulation, photosynthesis-related gene expression, antioxidant efficiency and redox status in wheat under mercury exposure.

Mercury (Hg) pollution is a global concern in cropland systems. Hg contamination causes a disruption in the growth, energy metabolism, redox balance, and photosynthetic activity of plants. In the removal of Hg toxicity, a recent critical strategy is the use of aerogels with biodegradability and biocompatibility. However, it is unknown how graphene oxide-based aerogels stimulate the defense systems in wheat plants exposed to Hg toxicity. Therefore, in this study, the photosynthetic, genetic, and biochemical effects of reduced graphene oxide aerogel treatments (gA; 50-100-250 mg L-1) were examined in wheat (Triticum aestivum) under Hg stress (50 µM HgCl2). The relative growth rate (RGR) significantly decreased (84%) in response to Hg stress. However, the reduced RGR and water relations (RWC) of wheat were improved by gA treatments. The impaired gas exchange levels (stomatal conductance, carbon assimilation rate, intercellular CO2 concentrations, and transpiration rate) caused by stress were reversed under Hg plus gAs. Additionally, stress hampered chlorophyll fluorescence (Fv/Fo, Fv/Fm), and under Hg toxicity the expression of psaA genes was reduced (>0.4-fold), but psaB gene was significantly up-regulated (>3-fold) which are the genes involved in PSI. By increasing expression patterns of both genes relating to PSI, gAs reversed the adverse consequences on Fv/Fo and Fv/Fm in the presence of excessive Hg concentration. The activities of glutathione S-transferase (GST), glutathione reductase (GR), catalase (CAT), ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR) decreased under Hg toxicity. On the other hand, the activities of superoxide dismutase (SOD), APX, GST, and glutathione peroxidase (GPX) increased following gA treatments against stress, leading to the successful elimination of toxic levels of H2O2 and lipid peroxidation (TBARS content) by decreasing the levels by about 30%, and 40%, respectively. By modulating enzyme/non-enzyme activity/contents including the AsA-GSH cycle, gAs contributed to the protection of the cellular redox state. Most important of all, gA applications were able to reduce Hg intake by approximately 66%. Therefore, these results showed that gAs were effective in highly inhibiting Hg uptake and could significantly increase wheat tolerance to toxicity by eliminating Hg-induced oxidative damage and inhibiting metabolic processes involved in photosynthesis. The findings obtained from the study provide a new perspective on the alleviation roles of reduced graphene oxide aerogels as an effective adsorbent for decreasing damages of mercury toxicity in wheat plants.

Polyamine, 1,3-diaminopropane, regulates defence responses on growth, gas exchange, PSII photochemistry and antioxidant system in wheat under arsenic toxicity.

The metalloid arsenic (As) is extremely hazardous to all living organisms, including plants. Pollution with As is very detrimental to the photosynthetic machinery, cell division, energy generation, and redox status. In order to cope with stress, the use of growth regulators such as polyamines (PA), which strengthen the antioxidant system of plants, has become widespread in recent years. PAs can modulate the plant growth through basic mechanisms common to all living organisms, such as membrane stabilization, free radical scavenging, deoxyribonucleic acid (DNA), ribonucleic acid (RNA) and protein synthesis, enzyme activities and second messengers. However, the effect of 1,3- diaminopropane (Dap), which is a product of PA catabolism, is not clear enough in plants exposed to As toxicity. In the current study, the different concentrations of 1,3-diaminopropane (0.1, 0.5 and 1 mM Dap) were hydroponically treated to wheat (Triticum aestivum) under arsenic stress (100 µM As) and then relative growth rate (RGR), relative water content (RWC), proline content (Pro), gas exchange parameters, PSII photochemistry, chlorophyll fluorescence kinetics, antioxidant activity and lipid peroxidation were assessed. RGR, RWC, osmotic potential and Pro content decreased in As-applied plants. The inhibition of these parameters could be reversed by Dap treatments. Besides, Dap applications mitigated the As toxicity-induced suppression on chlorophyll fluorescence (Fv/Fm, Fv/Fo and Fo/Fm) and the performance of PSII photochemistry. As impaired the balance on antioxidant capacity by decreased activities of catalase (CAT), peroxidase (POX), glutathione peroxidase (GPX), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and the contents of ascorbate (AsA) and glutathione (GSH) and then lipid peroxidation (TBARS content) increased. In the presence of Dap under As stress, the plants exhibited an increase in superoxide dismutase (SOD), POX, and GPX. Dap treatments contributed to the maintenance of cellular redox state (AsA/DHA and GSH/GSSG) by regulating the activities/contents of enzyme/non-enzyme involved in the AsA-GSH cycle. After Dap applications against stress, ROS accumulation (H2O2 content) and lipid peroxidation (TBARS) were effectively reduced. The findings showed that by eliminating As-induced oxidative damage and protecting the biochemical processes of photosynthesis, Dap treatments have a substantial potential to give resistance to wheat.

Protective role of quercetin and kaempferol against oxidative damage and photosynthesis inhibition in wheat chloroplasts under arsenic stress.

Arsenic (As) toxicity negatively impacts plant development, limits agricultural production, and, by entering the food chain, endangers human health. Studies on the use of natural and bioactive molecules in increasing plants' resistance to abiotic stressors, such as As, have gained increasing attention in the last few years. Flavonols are plant secondary metabolites with high potential in stress tolerance due to their roles in signal transmission. Therefore, the focus of this study was to examine the effects of two flavonols, quercetin (Q, 25 µM) and kaempferol (K, 25 µM), on growth parameters, photosynthesis, and chloroplastic antioxidant activity in wheat leaves under As stress (100 µM). As stress reduced the relative growth rate by 50% and relative water content by 25% in leaves. However, applying Q and/or K alleviated the As-induced suppression of growth and water relations. Exogenous phenolic treatments reversed the effects of As toxicity in photochemistry and maintained the photochemical quantum efficiency of the Photosystem II (Fv /Fm ). As exposure increased, the H2 O2 content in wheat chloroplasts by 42% and high levels of H2 O2 accumulation were also observed in guard cells in confocal microscopy images. Analysis of the chloroplastic antioxidant system has shown that Q and K applications increase the activity of antioxidant enzymes, including superoxide dismutase, peroxidase, and ascorbate peroxidase. Phenolic applications have induced the ascorbate-glutathione (AsA-GSH) cycle in charge of the protection of the cellular redox balance in different ways. It has been determined that Q triggers the AsA renewal, and K maintains the GSH pool. As a result, Q and K applications provide tolerance to wheat plants under As stress by increasing the chloroplastic antioxidant system activity and protecting photosynthetic reactions from oxidative damage. This study reveals the potential use of plant phenolic compounds in agricultural systems as a biosafe strategy to enhance plant stress tolerance, hence increasing yield.

Glutamate, Humic Acids and Their Combination Modulate the Phenolic Profile, Antioxidant Traits, and Enzyme-Inhibition Properties in Lettuce.

Lettuce (Lactuca sativa L., Asteraceae) is a popular vegetable leafy crop playing a relevant role in human nutrition. Nowadays, novel strategies are required to sustainably support plant growth and elicit the biosynthesis of bioactive molecules with functional roles in crops including lettuce. In this work, the polyphenolic profile of lettuce treated with glutamic acid (GA), humic acid (HA), and their combination (GA + HA) was investigated using an untargeted metabolomics phenolic profiling approach based on high-resolution mass spectrometry. Both aerial and root organ parts were considered, and a broad and diverse phenolic profile could be highlighted. The phenolic profile included flavonoids (anthocyanins, flavones, flavanols, and flavonols), phenolic acids (both hydroxycinnamics and hydroxybenzoics), low molecular weight phenolics (tyrosol equivalents), lignans and stilbenes. Overall, GA and HA treatments significantly modulated the biosynthesis of flavanols, lignans, low molecular weight phenolics, phenolic acids, and stilbene. Thereafter, antioxidant capacity was evaluated in vitro with 2,2-diphenyln-1-picrylhydrazyl (DPPH), 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP), and cupric ion reducing antioxidant capacity (CUPRAC) assays. In addition, this study examined the inhibitory properties of enzymes, including acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), tyrosinase, alpha-amylase, and alpha-glucosidase. Compared to individual treatments, the combination of GA + HA showed stronger antioxidant abilities in free radical scavenging and reducing power assays in root samples. Moreover, this combination positively influenced the inhibitory effects of root samples on AChE and BChE and the tyrosinase inhibitory effect of leaf samples. Concerning Pearson's correlations, antioxidant and enzyme inhibition activities were related to phenolic compounds, and lignans in particular correlated with radical scavenging activities. Overall, the tested elicitors could offer promising insights for enhancing the functional properties of lettuce in agricultural treatments.

Responses of individual and combined polystyrene and polymethyl methacrylate nanoplastics on hormonal content, fluorescence/photochemistry of chlorophylls and ROS scavenging capacity in Lemna minor under arsenic-induced oxidative stress.

Nanoplastics alter the adverse impacts of hazardous contaminants such as heavy metals by changing their adsorption and accumulation. Few findings are available on the interaction between nanoplastic and heavy metals in plants. However, there is no report on the mechanisms for removing metal stress-mediated oxidative damage by the combination treatments of nanoplastics. To address this lack of information, polystyrene nanoplastic (PS, 100 mg L-1) and polymethyl methacrylate (PMMA, 100 mg L-1) were hydroponically applied to Lemna minor exposed to arsenate (As, 100 µM) for 7 days. PS or PMMA caused a reduction in the contents of N, P, K, Ca, Mg and Mn, but the improved contents were detected in the presence of PS or PMMA plus As stress. The hormone contents (auxin, gibberellic acid, cytokinin, salicylic acid and jasmonic acid) reduced by stress were re-arranged through PS or PMMA applications. Based on chlorophyll efficiency, fluorescence kinetics and performance of PSII, the impaired photosynthesis by As stress was improved via PS or PMMA applications. This alleviation did not continue under the combined form of PS and PMMA in As-applied plants. All analyzed antioxidant activity (superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), glutathione reductase (GR), glutathione S-transferase (GST), glutathione peroxidase (GPX), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR)) decreased or unchanged under As, PS or PMMA. Due to the inactivation of the defense system, L. minor had high levels of hydrogen peroxide (H2O2) and thiobarbituric acid reactive substances (TBARS), showing lipid peroxidation. After As toxicity, induvial applications of PS or PMMA indicated the activated enzyme capacity (SOD, POX, GST and GPX) and upregulated AsA/DHA, GSH/GSSG and redox state of GSH, which facilitated the removal of radical accumulation. The efficiency of the antioxidant system in As + PS + PMMA-applied L. minor was not enough to remove damage induced by As stress; hereby, TBARS and H2O2 contents were similar to the As-treated group. Our findings from alone or combined application of PS and PMMA provide new information to advance the tolerance mechanism against As exposure in L. minor.

Hormetic activation of nano-sized rare earth element terbium on growth, PSII photochemistry, antioxidant status and phytohormone regulation in Lemnaminor.

Soils contaminated with rare earth elements (REEs) can damage agriculture by causing physiological disorders in plants which are evaluated as the main connection of the human food chain. A biphasic dose response with excitatory responses to low concentrations and inhibitory/harmful responses to high concentrations has been defined as hormesis. However, not much is clear about the ecological effects and potential risks of REEs to plants. For this purpose, here we showed the impacts of different concentrations of nano terbium (Tb) applications (5-10-25-50-100-250-500 mg L-1) on the accumulation of endogeneous certain ions and hormones, chlorophyll fluoresence, photochemical reaction capacity and antioxidant activity in duckweed (Lemna minor). Tb concentrations less than 100 mg L-1 increased the contents of nitrogen (N), phosphate (P), potassium (K+), calcium (Ca2+), magnesium (Mg2+), manganese (Mn2+) and iron (Fe2+). Chlorophyll fluorescence (Fv/Fm and Fv/Fo) was suppressed under 250-500 mg L-1 Tb. In addition, Tb toxicity affected the trapped energy adversely by the active reaction center of photosystem II (PSII) and led to accumulation of inactive reaction centers, thus lowering the detected level of electron transport from photosystem II (PSII) to photosystem I (PSI). On the other hand, 5-100 mg L-1 Tb enhanced the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), NADPH oxidase (NOX), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR) and glutathione S-transferase (GST). Tb (5-50 mg L-1) supported the maintenance of cellular redox status by promoting antioxidant pathways involved in the ascorbate-glutathione (AsA-GSH) cycle. In addition to the antioxidant system, the contents of some hormones such as indole-3-acetic acid (IAA), gibberellic acid (GA), cytokinin (CK) and salicylic acid (SA) were also induced in the presence of 5-100 mg L-1 Tb. In addition, the levels of hydrogen peroxide (H2O2) and lipid peroxidation (TBARS) were controlled through ascorbate (AsA) regeneration and effective hormonal modulation in L. minor. However, this induction in the antioxidant system and phytohormone contents could not be resumed after applications higher than 250 mg L-1 Tb. TBARS and H2O2, which indicate the level of lipid peroxidation, increased. The results in this study showed that Tb at appropriate concentrations has great potential to confer tolerance of duckweed by supporting the antioxidant system, protecting the biochemical reactions of photosystems and improving hormonal regulation.

Selectivity Tuning by Natural Deep Eutectic Solvents (NADESs) for Extraction of Bioactive Compounds from Cytinus hypocistis-Studies of Antioxidative, Enzyme-Inhibitive Properties and LC-MS Profiles.

In the present study, the extracts of Cytinus hypocistis (L.) L using both traditional solvents (hexane, ethyl acetate, dichloromethane, ethanol, ethanol/water, and water) and natural deep eutectic solvents (NADESs) were investigated in terms of their total polyphenolic contents and antioxidant and enzyme-inhibitive properties. The extracts were found to possess total phenolic and total flavonoid contents in the ranges of 26.47-186.13 mg GAE/g and 0.68-12.55 mg RE/g, respectively. Higher total phenolic contents were obtained for NADES extracts. Compositional differences were reported in relation to antioxidant potential studied by several assays (DPPH: 70.19-939.35 mg TE/g, ABTS: 172.56-4026.50 mg TE/g; CUPRAC: 97.41-1730.38 mg TE/g, FRAP: 84.11-1534.85 mg TE/g). Application of NADESs (choline chloride-urea 1:2, a so-called Reline) allowed one to obtain the highest number of extracts having antioxidant potential in the radical scavenging and reducing assays. NADES-B (protonated by HCl L-proline-xylitol 5:1) was the only extractant from the studied solvents that isolated a specific fraction without chelating activity. Reline extract exhibited the highest acetylcholinesterase inhibition compared to NADES-B and NADES-C (protonated by H2SO4 L-proline-xylitol 5:1) extracts, which showed no inhibition. The NADES extracts were observed to have higher tyrosinase inhibitory properties compared to extracts obtained by traditional organic solvents. Furthermore, the NADES extracts were relatively better inhibitors of the diabetic enzymes. These findings provided an interesting comparison in terms of total polyphenolic content yields, antioxidant and enzyme inhibitory properties (cholinesterase, amylase, glucosidase, and tyrosinase) between traditional solvent extracts and NADES extracts, used as an alternative. While the organic solvents showed better antioxidant activity, the NADES extracts were found to have some other improved properties, such as higher total phenolic content and enzyme-inhibiting properties, suggesting functional prospects for their use in phytonutrient extraction and fractionation. The obtained results could also be used to give a broad overview of the different biological potentials of C. hypocistis.

Multiwalled Carbon Nanotubes Alter the PSII Photochemistry, Photosystem-Related Gene Expressions, and Chloroplastic Antioxidant System in Zea mays under Copper Toxicity.

A critical approach against copper (Cu) toxicity is the use of carbon nanomaterials (CNMs). However, the effect of CNMs on Cu toxicity-exposed chloroplasts is not clear. The photosynthetic, genetic, and biochemical effects of multiwalled carbon nanotubes (50-100-250 mg L-1 CNT) were investigated under Cu stress (50-100 µM CuSO4) in Zea mays chloroplasts. Fv/Fm and Fv/Fo were suppressed under stress. Stress altered the antioxidant system and the expression of psaA, psaB, psbA, and psbD. The chloroplastic activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione S-transferase (GST), and glutathione peroxidase (GPX) increased under CNT + stress, and those of hydrogen peroxide (H2O2) and lipid peroxidation decreased. CNTs were promoted to the maintenance of the redox state by regulating enzyme/non-enzyme activity/contents involved in the AsA-GSH cycle. Furthermore, CNTs inverted the negative effects of Cu by upregulating the transcriptions of photosystem-related genes. However, the high CNT concentration had adverse effects on the antioxidant capacity. CNT has great potential to confer tolerance by reducing Cu-induced damage and protecting the biochemical reactions of photosynthesis.

Fe2O3-modified graphene oxide mitigates nanoplastic toxicity via regulating gas exchange, photosynthesis, and antioxidant system in Triticum aestivum.

The ever-increasing plastic pollution in soil and water resources raises concerns about its effects on terrestrial plants and agroecosystems. Although there are many reports about the contamination with nanoplastics on plants, the presence of magneto-assisted nanomaterials enabling the removal of their adverse impacts still remains unclear. Therefore, the purpose of the current study is to evaluate the potential of nanomaterial Fe2O3-modified graphene oxide (FGO, 50-250 mg L-1) to eliminate the adverse effects of nanoplastics in plants. Wheat plants exposed to polystyrene nanoplastics concentrations (PS, 10, 50 and 100 mg L-1) showed decreased growth, water content and loss of photosynthetic efficiency. PS toxicity negatively altered gas exchange, antenna structure and electron transport in photosystems. Although the antioxidant system was partially activated (only superoxide dismutase (SOD), NADPH oxidase (NOX) and glutathione reductase (GR)) in plants treated with PS, it failed to prevent PS-triggered oxidative damage, as showing lipid peroxidation and hydrogen peroxide (H2O2) levels. FGOs eliminated the adverse impacts of PS pollution on growth, water status, gas exchange and oxidative stress markers. In addition, FGOs preserve the biochemical reactions of photosynthesis by actively increasing chlorophyll fluorescence parameters in the stressed-wheat leaves. The activities of all enzymatic antioxidants increased, and the H2O2 and TBARS contents decreased. GSH-mediated detoxifying antioxidants such as glutathione S-transferase (GST) and glutathione peroxidase (GPX) were stimulated by FGOs against PS pollution. FGOs also triggered the enzymes and non-enzymes related to the Asada-Halliwell cycle and protected the regeneration of ascorbate (AsA) and glutathione (GSH). Our findings indicated that FGO had the potential to mitigate nanoplastic-induced damage in wheat by regulating water relations, protecting photosynthesis reactions and providing efficient ROS scavenging with high antioxidant capacity. This is the first report on removing PS-induced damage by FGO applications in wheat leaves.

Polystyrene nanoplastic contamination mixed with polycyclic aromatic hydrocarbons: Alleviation on gas exchange, water management, chlorophyll fluorescence and antioxidant capacity in wheat.

Polycyclic aromatic hydrocarbons (PAHs) constitute a significant environmental pollution group that reaches toxic levels with anthropogenic activities. The adverse effects of nanoplastics accumulating in ecosystems with the degradation of plastic wastes are also a growing concern. Previous studies have generally focused on the impact of single PAH or plastic fragments exposure on plants. However, it is well recognized that these contaminants co-exist at varying rates in agricultural soil and water resources. Therefore, it is critical to elucidate the phytotoxicity and interaction mechanisms of mixed pollutants. The current study was designed to comparatively investigate the single and combined effects of anthracene (ANT, 100 mg L-1), fluorene (FLU, 100 mg L-1) and polystyrene nanoplastics (PS, 100 mg L-1) contaminations in wheat. Plants exposed to single ANT, FLU and PS treatments demonstrated decline in growth, water content, high stomatal limitations and oxidative damage. The effect of ANT + FLU on these parameters was more detrimental. In addition, ANT and/or FLU treatments significantly suppressed photosynthetic capacity as determined by carbon assimilation rate (A) and chlorophyll a fluorescence transient. The antioxidant system was not fully activated (decreased superoxide dismutase, peroxidase and glutathione reductase) under ANT + FLU, then hydrogen peroxide (H2O2) content (by 2.7-fold) and thiobarbituric acid reactive substances (TBARS) (by 2.8-fold) increased. Interestingly, ANT + PS and FLU + PS improved the growth, water relations and gas exchange parameters. The presence of nanoplastics recovered the adverse effects of ANT and FLU on growth by protecting the photosynthetic photochemistry and reducing oxidative stress. PAH plus PS reduced the ANT and FLU accumulation in wheat leaves. In parallel, the increased antioxidant system, regeneration of ascorbate, glutathione and glutathione redox status observed under ANT + PS and FLU + PS. These findings will provide an information about the phytotoxicity mechanisms of mixed pollutants in the environment.

An In-Depth Study on the Metabolite Profile and Biological Properties of Primula auriculata Extracts: A Fascinating Sparkle on the Way from Nature to Functional Applications.

The biological activity of the aerial part and rhizomes of Primula auriculata were assessed for the first time. The biological activities (antioxidant properties, enzyme inhibition, and AGE inhibition) as well as the phenolic and flavonoid contents of the ethyl acetate, ethanol, hydro-ethanol and water extracts of P. auriculata aerial parts and rhizomes were determined. Cell viability assays and gelatin zymography were also performed for MMP-2/-9 to determine the molecular mechanisms of action. The gene expression for MMPs was described with RT-PCR. The levels of various proteins, including phospho-Nf-κB, BCL-2, BAX, p-53, and cyclin D1 as well as RAGE were measured using Western blot analysis. The hydro-ethanol extract of the aerial part possessed the highest phenolic (56.81 mg GAE/g) and flavonoid (63.92 mg RE/g) contents. In-depth profiling of the specialized metabolites by ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) allowed for the identification and annotation of 65 compounds, including phenolic acids and glycosides, flavones, flavonols, chalcones, dihydrochalcones, and saponins. The hydro-ethanol extract of the aerial parts (132.65, 180.87, 172.46, and 108.37 mg TE/g, for the DPPH, ABTS, CUPRAC, and FRAP assays, respectively) and the ethanol extract of the rhizomes (415.06, 638.30, 477.77, and 301.02 mg TE/g, for the DPPH, ABTS, CUPRAC, and FRAP assays, respectively) exhibited the highest free radical scavenging and reducing activities. The ethanol and hydro-ethanol extracts of both the P. auriculata aerial part and rhizomes exhibited higher inhibitory activity against acetylcholinesterase, while the hydro-ethanol extracts (1.16 mmol ACAE/g, for both the aerial part and rhizomes extracts) were more active in the inhibition of α-glucosidase. After the treatment of an HT-29 colorectal cancer cell line with the extracts, the apoptosis mechanism was initiated, the integrity of the ECM was remodeled, and cell proliferation was also taken under control. In this way, Primula extracts were shown to be potential drug sources in the treatment of colorectal cancer. They were also detected as natural MMP inhibitors. The findings presented in the present study appraise the bioactivity of P. auriculata, an understudied species. Additional assessment is required to evaluate the cytotoxicity of P. auriculata as well as its activity in ex vivo systems.

The hormetic dose-risks of polymethyl methacrylate nanoplastics on chlorophyll a fluorescence transient, lipid composition and antioxidant system in Lactuca sativa.

Nanoplastic pollution has become an increasing problem due to over-consumption and degradation in ecosystems. A little is known about ecological toxicity and the potential risks of nanoplastics on plants. To better comprehend the hormetic effects of nanoplastics, the experimental design was conducted on the impacts of polymethyl methacrylate (PMMA) on water status, growth, gas exchange, chlorophyll a fluorescence transient, reactive oxygen species (ROS) content (both content and fluorescence visualization), lipid peroxidation and antioxidant capacity (comparatively between leaves and roots). For this purpose, PMMA (10, 20, 50 and 100 mg L-1) was hydroponically applied to Lactuca sativa for 15 days(d). PMMA exposure resulted a decline in the growth, water content and osmotic potential. As based on assimilation rate (A), stomatal conductance (gs), and intercellular CO2 concentrations (Ci), the decreased stomatal limitation (Ls) and, A/Ci and increased intrinsic mesophyll efficiency proved low carboxylation efficiency showing impaired photosynthesis as a non-stomatal limitation. PMMA toxicity increased the trapping fluxes and absorption with a decrease in electron transport fluxes caused the disruption in reaction centers of photosystems. The leaves and roots had a similar effect against PMMA toxicity, with increased superoxide dismutase (SOD) activity. Although, catalase (CAT) and peroxidase (POX) of leaves increased under 10 mg L-1 PMMA, these defense activities failed to prevent radicals from attacking. Compared to the leaves, the lettuce roots showed an intriguing result for AsA-GSH cycle against PMMA exposure. In the roots, the lowest PMMA application provided the high ascorbate/dehydroascorbate (AsA/DHA), GSH/GSSG and the pool of AsA/glutathione (GSH) and non-suppressed GSH redox state. Also, 10 mg L-1 PMMA helped remove high hydrogen peroxide (H2O2) by both glutathione peroxidase (GPX) and glutathione S-transferase (GST). Since this improvement in the antioxidant system could not be continued in roots after higher applications than 20 mg L-1 PMMA, TBARS (Thiobarbituric acid-reactive substances), indicating the level of lipid peroxidation, and H2O2 increased. Our findings obtained from PMMA-applied lettuce provide new information to advance the tolerance mechanism against nanoplastic pollution.

The effects of fullerene on photosynthetic apparatus, chloroplast-encoded gene expression, and nitrogen assimilation in Zea mays under cobalt stress.

Carbon nanostructures, such as the water-soluble fullerene (FLN) derivatives, are considered perspective agents for agriculture. FLN can be a novel nano-agent modulating plant response against stress conditions. However, the mechanism underlying the impacts of FLN on plants in agroecosystems remains unclear. Zea mays was exposed to exogenous C60 -FLN applications (FLN1: 100; FLN2: 250; and FLN3: 500 mg L-1 ) with/without cobalt stress (Co, 300 µM) for 3 days (d). In the maize chloroplasts, Co stress disrupted the photosynthetic efficiency and the expression of genes related to the photosystems (psaA and psbA). FLNs effectively improved the efficiency and photochemical reaction of photosystems. Co stress induced the accumulation of reactive oxygen species (ROS) as confirmed by ROS-specific fluorescence in guard cells. Co stress increased only chloroplastic superoxide dismutase (SOD) and peroxidase (POX). Stress triggered oxidative damages in maize chloroplasts, measured as an increase in TBARS content. In Co-stressed seedlings exposed to FLN1 and FLN2 exposures, the hydrogen peroxide (H2 O2 ) was scavenged through the nonenzymes/enzymes-related to the AsA-GSH cycle by preserving ascorbate (AsA) conversion, as well as GSH/GSSG and glutathione (GSH) redox state. Also, the alleviation effect of FLN3 against stress could be attributed to increased glutathione S-transferase (GST) activity and AsA regeneration. FLN applications reversed the inhibitory effects of Co stress on nitrogen assimilation. In maize chloroplasts, FLN increased the activities of nitrate reductase (NR), glutamate dehydrogenase (GDH), nitrite reductase (NiR), and glutamine synthetase (GS), which provided conversion of inorganic nitrogen (N) into organic N. The ammonium (NH4 + ) toxicity was removed via GS and GDH but not glutamate synthase (GOGAT). The increased NAD-GDH (deaminating) and NADH-GDH (aminating) activities indicated that GDH was needed more for NH4 + detoxification. Therefore, FLN exposure to Co-stressed maize plants might play a role in N metabolism regarding the partitioning of N assimilates. Exogenous FLN conceivably removed Co toxicity by improving the expressions of genes related to reaction center proteins of photosystems, increasing the level of enzymes related to the defense system, and improving the N assimilation in maize chloroplasts.

Untargeted Phenolic Profiling and Functional Insights of the Aerial Parts and Bulbs of Drimia maritima (L.) Stearn.

Drimia maritima (L.) Stearn (squill), belonging to the Asparagaceae family, is acknowledged as a medicinally valuable species from the Drimia genera. In this study, water, methanol, and ethyl acetate extracts of D. maritima aerial parts and bulbs were investigated for their polyphenols profile and evaluated for their antioxidant and enzyme inhibition properties. Phenolics were profiled through an untargeted metabolomics approach using an ultra-high pressure liquid chromatograph coupled to quadrupole-time-of-flight mass spectrometry (UHPLC-QTOF-MS). This analysis revealed an enrichment of low molecular weight phenolics and flavonoids in the aerial parts of D. maritima, while lignans mainly characterized bulb extracts. Antioxidant capacity was investigated by different assays, including phosphomolybdenum assays, radical scavenging (DPPH: 2,2-diphenyl-1-picrylhydrazyl; ABTS: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), as well as reducing ability (CUPRAC: cupric reducing antioxidant capacity; FRAP: ferric reducing antioxidant power), and metal chelating. In radical scavenging and reducing power assays, the water extract of aerial parts exhibited the strongest ability (DPPH: 36.99 mg trolox equivalent (TE)/g; ABTS: 85.96 mg TE/g; CUPRAC: 87.37 mg TE/g; FRAP: 55.43 mg TE/g). In general, the ethyl acetate extracts from aerial parts and bulbs provided the weakest antioxidant capacity. Concerning enzyme inhibitory activities, the water extracts of the bulb were poorly active, while the ethyl acetate extracts from both plant portions displayed the best α-amylase inhibitory abilities. The best acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) abilities were recorded by ethyl acetate extract of aerial parts (2.36 mg galantamine equivalent (GALAE)/g) and bulbs (5.10 mg GALAE/g), respectively. Overall, these results support the medicinal aptitude of D. maritima and its possible use as a natural source of antioxidants and enzyme inhibitors with functional potential.