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In tissue culture, efficient nutrient availability and effective control of callus contamination are crucial for successful plantlet regeneration. This study was aimed to enhance callogenesis, callus regeneration, control callus contamination, and substitute iron (Fe) source with FeO-NPs in Murashige and Skoog (MS) media. Nanogreen iron oxide (FeO-NPs) were synthesized and well characterized with sizes ranging from 2 to 7.5 nm. FeO-NPs as a supplement in MS media at 15 ppm, significantly controlled callus contamination by (80%). Results indicated that FeCl 3 -based FeO-NPs induced fast callus induction (72%) and regeneration (43%), in contrast FeSO 4 -based FeO-NPs resulted in increased callus weight (516%), diameter (300%), number of shoots (200%), and roots (114%). Modified media with FeO-NPs as the Fe source induced fast callogenesis and regeneration compared to normal MS media. FeO-NPs, when applied foliar spray, increased Plant fresh biomass by 133% and spike weight by 350%. Plant height increased by 54% and 33%, the number of spikes by 50% and 265%, and Chlorophyll content by 51% and 34% in IRRI-6 and Kissan Basmati, respectively. Additionally, APX (Ascorbate peroxidase), SOD (Superoxide dismutase), POD (peroxidase), and CAT (catalase) increased in IRRI-6 by 27%, 29%, 283%, 62%, while in Kissan Basmati, APX increased by 70%, SOD decreased by 28%, and POD and CAT increased by 89% and 98%, respectively. Finally, FeO-NPs effectively substituted Fe source in MS media, shorten the plant life cycle, and increase chlorophyll content as well as APX, SOD, POD, and CAT activities. This protocol is applicable for tissue culture in other cereal crops as well.

Bioactive compounds are secondary metabolites of plants. They offer diverse pharmacological properties. Peganum harmala is reported to have pharmaceutical effects like insecticidal, antitumor, curing malaria, anti-spasmodic, vasorelaxant, antihistaminic effect. Rosa brunonii has medicinal importance in its flower and fruits effective against different diseases and juice of leaf is reported to be applied externally to cure wounds and cuts. Dryopteris ramosa aqueous leaf extract is used to treat stomach ulcers and stomachaches. Each of these three medicinal plants have been indicated to have anticancer, antiviral, antioxidant, cytotoxic and antifungal effects but efficacy of their bioactive compounds remained unexplored. Study was aimed to explore In-vitro and In-silico anticancer, antiviral, antioxidant, cytotoxic and antifungal effects of bioactive compounds of above three medicinal plants. DPPH and ABTS assay were applied for assessment of antioxidant properties of compounds. Antibacterial properties of compounds were checked by agar well diffusion method. Brine shrimp lethality assay was performed to check cytotoxic effect of compounds. Molecular docking was conducted to investigate the binding efficacy between isolated compounds and targeted proteins. The compound isomangiferrin and tiliroside presented strong antioxidant potential 78.32% (± 0.213) and 77.77% (± 0.211) respectively in DPPH assay while harmaline showed 80.71% (± 0.072) at 200 µg/mL in ABTS assay. The compound harmine, harmaline and PH-HM 17 exhibited highest zone of inhibition 22 mm, 23 mm, 22 mm respectively against Xanthomonas while Irriflophenone-3-C-β- D-glucopyranoside showed maximum zone of inhibition 34 mm against E. coli. The compound isomangiferrin and vasicine contained strong antibacterial activity 32 mm and 22 mm respectively against S. aureus. The compound mangiferrin, astragalin, tiliroside, quercitin-3-O-rhamnoside showed maximum inhibitory zone 32 mm, 26 mm, 24 mm and 22 mm respectively against Klebsiella pneumoniae. Highest cytotoxic effect was observed by compound tiliroside i.e. 95% with LD 50 value 73.59 µg/mL. The compound tiliroside showed the best binding mode of interaction to all targeted proteins presenting maximum hydrophobic interactions and hydrogen bonds. The binding affinity of tiliroside was − 17.9, − 14.9, − 14.6, − 13.8, − 12.8 against different proteins 6VAR, 5C5S, IEA3, 2XV7 and 6LUS respectively. Bioactive compounds are significant natural antioxidants, which could help to prevent the progression of various diseases caused by free radicals. Based on molecular docking we have concluded that phytochemicals can have better anticancer and antiviral potential.

Advent of proteomic techniques has made it possible to identify a broad spectrum of proteins in living systems. Studying the impact of nanoparticle (NP)-mediated plant protein responses is an emerging field. NPs are continuously being released into the environment and directly or indirectly affect plant’s biochemistry. Exposure of plants to NPs, especially crops, poses a significant risk to the food chain, leading to changes in underlying metabolic processes. Once absorbed by plants, NPs interact with cellular proteins, thereby inducing changes in plant protein patterns. Based on the reactivity, properties, and translocation of nanoparticles, NPs can interfere with proteins involved in various cellular processes in plants such as energy regulation, redox metabolism, and cytotoxicity. Such interactions of NPs at the subcellular level enhance ROS scavenging activity, especially under stress conditions. Although higher concentrations of NPs induce ROS production and hinder oxidative mechanisms under stress conditions, NPs also mediate metabolic changes from fermentation to normal cellular processes. Although there has been lots of work conducted to understand the different effects of NPs on plants, the knowledge of proteomic responses of plants toward NPs is still very limited. This review has focused on the multi-omic analysis of NP interaction mechanisms with crop plants mainly centering on the proteomic perspective in response to both stress and non-stressed conditions. Furthermore, NP-specific interaction mechanisms with the biological pathways are discussed in detail.

Heavy metal (HM) contamination of soil due to anthropogenic activities has led to bioaccumulation and biomagnification, posing toxic effects on plants by interacting with vital cellular biomolecules such as DNA and proteins. Brassica species have developed complex physiological, biochemical, and molecular mechanisms for adaptability, tolerance, and survival under these conditions. This review summarizes the HM tolerance strategies of Brassica species, covering the role of root exudates, microorganisms, cell walls, cell membranes, and organelle-specific proteins. The first line of defence against HM stress in Brassica species is the avoidance strategy, which involves metal ion precipitation, root sorption, and metal exclusion. The use of plant growth-promoting microbes, Pseudomonas , Psychrobacter , and Rhizobium species effectively immobilizes HMs and reduces their uptake by Brassica roots. The roots of Brassica species efficiently detoxify metals, particularly by flavonoid glycoside exudation. The composition of the cell wall and callose deposition also plays a crucial role in enhancing HMs resistance in Brassica species. Furthermore, plasma membrane–associated transporters, BjCET , BjPCR , BjYSL , and BnMTP , reduce HM concentration by stimulating the efflux mechanism. Brassica species also respond to stress by up-regulating existing protein pools or synthesizing novel proteins associated with HM stress tolerance. This review provides new insights into the HM tolerance mechanisms of Brassica species, which are necessary for future development of HM-resistant crops.

Brassica species, capable of heavy metals (HMs) hyperaccumulation, differ in their ability to accumulate and tolerate metals present in their environment. In this comparative study, the accumulation, morphological, and physiological responses of three Brassica species i.e., Brassica juncea, B. napus, and B. campestris , against lead (Pb) were examined. Plants were grown in pots under greenhouse conditions and subjected to 0, 50, 100, 150 mM concentrations of Pb for 14 days. The study revealed that 150 mM Pb concentration reduced the plant length and biomass in all the species and this decline was more obvious in B. napus . At 100 mM Pb concentration, plant length increased 3.5% in B. juncea , while decreased by 8 and 36% in B. campestris and B. napus , respectively. B. campestris and B. napus suffered from more pronounced Pb-accumulation in the root followed by shoot as compared to B. juncea . Pb-accumulation in 100 mM treated root of B. campestris and B. napus increased 29 and 80%, respectively as compared to B. juncea Pb treated root. Antioxidant enzyme catalase (CAT) activity was increased in B. juncea and B. campestris up to 150 mM concentration, while in B. napus activity of enzyme decreased at 100 and 150 mM Pb concentration. Phenylalanine ammonia-lyase (PAL) and nitrate reductase activity increased at 50 mM, while the polyphenol oxidase (PPO) and nitrite reductase significantly increased at 150 mM. Brassica species also showed more significant accumulation of amino acid, inhibition of proteins and total sugar content at 100 and 150 mM concentrations. Although all species exhibited enhanced antioxidant activity, activation in B. juncea was relatively higher. These results suggest that B. juncea is relatively more tolerant towards Pb stress as compared to B. campestris and B. napus due to reduced metal uptake and enhanced antioxidant enzyme activities.

Lead (Pb) accumulation, even in minute quantities, has adverse effects on the morphology, physiology, and biochemistry of almost all plants, resulting in various abnormalities. Silicon dioxide nanoparticles (SiO 2 -NPs) are used excessively to reduce abiotic stresses in a large variety of plant species. The present research work was designed to explore the role of SiO 2 -NPs in the mitigation of Pb toxicity in Ocimum basilicum . SiO 2 -NPs were green-synthesized from Arando donax plant extract. Characterization of green synthesized SiO 2 -NPs was assessed with UV-vs, XRD, FTIR, and SEM–EDS. To analyze the morphology and antioxidant enzyme activities in O. basilicum , 8 days old plants were subjected to 3 different concentrations of Pb and SiO 2 -NPs (50, 500, and 1000 ppm). Results of UV-vs, XRD, FTIR, and SEM–EDS showed the capping of SiO 2- NPs by different functional groups (Si (CH 3 ) 3 , and Si–O-Si) together with its crystalline structure. The average size of the nanoparticles was 26 nm which was confirmed by XRD analysis. Morphological analysis revealed that treatment with 500 ppm concentration of Pb resulted in a significant decrease in the length of root, shoot, and weight, in the ratio of 19, 14, and 10%, respectively. But treatment with 500 ppm (SiO 2 - NPs) significantly promoted root, shoot length, and weight of the plant, at the rate of 13, 22, and 7%, respectively. After the confirmation of ameliorative effect of SiO 2 -NPs, combined application of Pb + SiO 2 -NPs was tested. Root damage and Pb concentration in all the plant parts were much reduced. It was revealed that antioxidant activities of POD and APX were markedly decreased while those of the CAT and SOD increased. Results revealed that SiO 2 -NPs are an anti-stressor, that removes Pb from O. basilicum , by enhancing its antioxidant activity. Graphical Abstract

Diabetes is a chronic metabolic disorder that affects an increasing number of people worldwide, frequently managed with synthetic drugs that have side effects and can be costly. Apteranthes tuberculata (N.E.Br.) Meve & Liede, a plant with traditional medicinal use in Pakistan to treat diabetes, but its antidiabetic potential has not been scientifically validated. This research assessed the phytochemicals, antioxidant properties, and dipeptidyl peptidase‐4 (DPP‐4) inhibitory activity of A. tuberculata's methanolic extract. The extract was assessed through in vitro antioxidant assays, DPP‐4 inhibition test, and metabolomic analysis via Fourier‐transform infrared (FTIR) spectroscopy and liquid chromatography–mass spectrometry (LC–MS). The study used computational tools to visualize compound structures, protein‐ligand interactions, and to measure pharmacokinetic parameters. Phytochemical analysis revealed significant levels of total phenols (71.991 ± 0.78 mg/g gallic acid equivalents) and flavonoids (66.216 ± 0.09 mg/g quercetin equivalents). Results showed a robust total antioxidant capacity (70.900 ± 2 mg/g ascorbic acid), total reducing power (72.000 ± 2.00 mg/g gallic acid equivalents), and DPPH IC50 value of 96.54 μg/mL. FTIR spectra showed the presence of carbohydrates and glycosides. The extract exhibited 70% DPP‐4 inhibitory activity (IC50 value = 46.761 ± 0.043 μg/mL), comparable to Sitagliptin at 78% (IC50 value = 20.474 ± 0.407 μg/mL). LC–MS identified 24 bioactive compounds, including flavonoids and glycosides, with compounds like Kaempferol‐3‐O‐rutinoside‐7‐O‐glucoside and Kaempferol‐7‐O‐rutinoside showing strong binding interactions with DPP‐4. These results underscore the therapeutic potential of A. tuberculata as a natural source of DPP‐4 inhibitors for managing diabetes. Metabolomics revealed 24 major compounds, mainly glycosides, flavonoids, and triterpenes. Antidiabetic compounds Kaempferol‐7‐O‐rutinoside and Kaempferol‐3‐O‐rutinoside‐7‐O‐glucoside show maximum inhibiting activity for DPP4 protein (in vitro and in silico). Out of 24 compounds, 5 demonstrate no PAINS and break alerts.

Diabetes is a chronic metabolic disorder that affects an increasing number of people worldwide, frequently managed with synthetic drugs that have side effects and can be costly. Apteranthes tuberculata ( N.E.Br .) Meve & Liede, a plant with traditional medicinal use in Pakistan to treat diabetes, but its antidiabetic potential has not been scientifically validated. This research assessed the phytochemicals, antioxidant properties, and dipeptidyl peptidase‐4 (DPP‐4) inhibitory activity of A. tuberculata 's methanolic extract. The extract was assessed through in vitro antioxidant assays, DPP‐4 inhibition test, and metabolomic analysis via Fourier‐transform infrared (FTIR) spectroscopy and liquid chromatography–mass spectrometry (LC–MS). The study used computational tools to visualize compound structures, protein‐ligand interactions, and to measure pharmacokinetic parameters. Phytochemical analysis revealed significant levels of total phenols (71.991 ± 0.78 mg/g gallic acid equivalents) and flavonoids (66.216 ± 0.09 mg/g quercetin equivalents). Results showed a robust total antioxidant capacity (70.900 ± 2 mg/g ascorbic acid), total reducing power (72.000 ± 2.00 mg/g gallic acid equivalents), and DPPH IC 50 value of 96.54 μg/mL. FTIR spectra showed the presence of carbohydrates and glycosides. The extract exhibited 70% DPP‐4 inhibitory activity (IC 50 value = 46.761 ± 0.043 μg/mL), comparable to Sitagliptin at 78% (IC 50 value = 20.474 ± 0.407 μg/mL). LC–MS identified 24 bioactive compounds, including flavonoids and glycosides, with compounds like Kaempferol‐3‐O‐rutinoside‐7‐O‐glucoside and Kaempferol‐7‐O‐rutinoside showing strong binding interactions with DPP‐4. These results underscore the therapeutic potential of A. tuberculata as a natural source of DPP‐4 inhibitors for managing diabetes.

Chemical quality and extent of pollution in effluent-contaminated irrigation water collected from two sectors (X and Y) of peri-urban southern regions of Khyber Pakhtunkhwa, Pakistan, were investigated. Various physico-chemical parameters like pH, EC, TS, TSS, TDS, TH, cations (Na 1+ , K 1+ , Li 1+ , Ca 2+ , Mg 2+ ), anions (Cl 1− , F 1− , HCO 3 1− , NO 3 1− , SO 4 2− ), and heavy metals (Cr, Fe, Cu, Zn, Pb, Cd) were determined. The data indicated wide ranges of variation in all the parameters. Concentration of physico-chemical properties such as EC, TS, TSS, TH, Li 1+ , Ca 2+ , Mg 2+ , and HCO 3 1− and heavy metals (Cr, Fe, Cu, and Cd) exceeded WHO permissible limits. The distribution of heavy metals in two sampling areas followed the increasing order as Fe > Cr > Cd > Cu > Pb > Zn. The Piper diagram showed that water in the region was of alkaline type. Strong positive correlations were observed among most of the parameters; however, pH was negatively correlated with most of the parameters. According to principle component analysis, parameters causing variations in water quality were mainly EC, TDS, TS, TSS, TH, Na 1+ , K 1+ , Ca 2+ , Mg 2+ , F 1− , NO 3 1− , SO 4 2− , Cr, Fe, Cu, and Zn. Linear regression analysis revealed that heavy metals like Cr, Fe, and Cu concentrations were strongly correlated having an R 2 value of 0.92 at P  ≤ 0.001 with pH. Overall, the results suggested that irrigation water of the area was not fit for irrigation purposes and might cause serious threats for safe, healthy food and feed production. Proper monitoring of water of the said area should be carried out, and quality of irrigation water should be checked from time to time.

The impact of city effluents on water quality of Indus River was assessed in the southern region of Khyber Pakhtunkhwa, Pakistan. Water samples were collected in dry ( DS ) and wet ( WS ) seasons from seven sampling zones along Indus River and the physical, bacteriological, and chemical parameters determining water quality were quantified. There were marked temporal and spatial variations in the water quality of Indus River. The magnitude of pollution was high in WS compared with DS . The quality of water varied across the sampling zones, and it greatly depended upon the nature of effluents entering the river. Water samples exceeded the WHO permissible limits for pH, EC, TDS, TS, TSS, TH, DO, BOD, COD, total coliforms, Escherichia coli , Ca 2+ , Mg 2+ , NO 3 − , and PO 4 2− . Piper analysis indicated that water across the seven sampling zones along Indus River was alkaline in nature. Correlation analyses indicated that EC, TDS, TS, TH, DO, BOD, and COD may be considered as key physical parameters, while Na + , K + , Ca 2+ , Mg 2+ , Cl − , F − , NO 3 − , PO 4 2− , and SO 4 2− as key chemical parameters determining water quality, because they were strongly correlated ( r  > 0.70) with most of the parameters studied. Cluster analysis indicated that discharge point at Shami Road is the major source of pollution impairing water quality of Indus River. Wastewater treatment plants must be installed at all discharge points along Indus River for protecting the quality of water of this rich freshwater resource in Pakistan.