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Juniperus procera is a natural source of bioactive compounds with the potential of antitumor, antimicrobial, insecticidal, antifungal, and antioxidant activities. An optimization method was developed for total phenolic content (TPC), total flavonoid content (TFC), and total tannin content (TTC) in leaf and seed extract of Juniperus procera. Organic solvents (methanol (99.8%), ethanol (99%), and acetone (99.5%)), and deionized water (DI) were used for extraction. The estimation of TPC, TFC, and TTC in plant materials was carried out using UV-spectrophotometer and HPLC with the standards gallic acid, quercetin, and tannic acid. Recovery of TPC in leaf extract ranged from 2.9 to 9.7 mg GAE/g DW, TFC from 0.9 to 5.9 mg QE/g DW, and TTC ranged from 1.5 to 4.3 mg TA/g DW while the TPC value in the seed extract ranged from 0.53 to 2.6 mg GAE/g DW, TFC from 0.5 to 1.6 mg QE/g DW, and TTC ranged from 0.5 to 1.4 mg TA/g DW. This result revealed that methanol is the best solvent for recovery of the TPC value (9.7 mg) from leaf extract in comparison to other solvents. Ethanol recorded the highest result of TFC (5.9 mg) in leaf extract among the solvents whereas acetone was the best for TTC yield recovery from leaf extract (4.3 mg). In the case of the seed extract, ethanol was the best solvent for both TPC (2.6 mg), and TFC (1.6 mg) recovery in comparison to other solvents. Total tannin content in methanol resulted in significant recovery from seed extract (1.4 mg). Separation and quantification of gallic acid, quercetin, and tannic acid in plant materials were undertaken using HPLC. Gallic acid in leaf and seed of J. procera ranged from 6.6 to 9.2, 6.5 to 7.2 µg/g DW, quercetin from 6.3 to 18.2, 0.9 to 4.2 µg/g DW, and tannic acid from 16.2 to 29.3, 6.6 to 9.3 µg/g DW, respectively. Solvents have shown a significant effect in the extraction of phenolic compounds. Moreover, phytochemicals in plant materials were identified using GC-MS and resulted in very important bioactive compounds, which include anti-inflammatory, antibacterial, and antitumor agents such as ferruginol, phenanthrene, and n-hexadecanoic acid. In conclusion, the optimal solvent for extraction depends on the part of the plant material and the compounds that are to be isolated.

Biosynthesized nanoparticles have played vital role recently, as suggested to be alternative to physical and chemical methods. In this study, biosynthesis of zinc oxide nanoparticles (ZnO NPs) were carried out using leaf extracts of Phoenix dactylifera L. and Zinc nitrate. The effect of ZnO nanoparticles on biomass and biochemical parameters was investigated. Biosynthesized ZnO nanostructure was characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM), UV–visible spectrophotometer and Fourier transform infrared spectroscopy (FTIR). Which resulted in spherical shape with size ranging between 16 to 35 nm of Biosynthesized ZnO nanoparticles and UV absorption beak at 370.5 nm with clear peaks of functional groups. The impact of different concentrations (0.0 mg/L, 80 mg/L and 160 mg/L) of biosynthesized ZnO nanoparticles on biomass and bioactive compounds production of Juniperus procera in vitro was investigated. The results showed that, biosynthesized ZnO NPs (80 mg/L and 160 mg/L) concentrations were boosted the growth of J. Procera with significantly compared to non-treated plants in vitro. The highest concentration (160 mg/L) of ZnO NPs was enhanced the growth of plant at beginning period, one month later shoots became yellow and callus turned to be brownish. Moreover, the influence of ZnO NPs on phytochemical compounds in callus of Juniperus procera was examined using GC–MS analysis. The differences among treatments were recoded . Overall, zinc oxide nanoparticles substantially improved the growth of shoots and callus with increasing of biochemical parameters such as chlorophyll a, total phenolic and flavonoids contents, besides the total protein and, SOD, CAT and APX activity. ZnO NPs might be induced some phytochemical compounds as well as inhibit.

Bioactive compounds of medicinal plants present as natural ingredients provide health benefits beyond the basic nutritional value of these products. However, the availability of bioactive compounds in the current natural sources is limited. Hence, the induction of bioactive compound production from medicinal plants through nanoparticles (NPs) might play a vital role in industrially important medicinal compounds. Therefore, this study aimed to synthesize silver nanoparticles (AgNPs) biologically and to investigate their effect on phytochemical compound production from the callus of Juniperus procera . AgNPs were synthesized biologically using aqueous leaf extract of Phoenix dactylifera , which acted as a reducing and capping agent, and silver nitrate solution. The formation of AgNPs has been confirmed through different analytical techniques such as UV-Visible spectroscopy (UV), Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), and scanning electron microscope (SEM). The impact of different concentrations (0.0, 5, 20, and 50 mg/L) of AgNPs on enzymatic and non-enzymatic antioxidants of the callus of J. procera was investigated. The obtained results showed a significant effect of AgNPs on biomass accumulation and non-enzymatic antioxidants (phenol, tannin, and flavonoid content). Additionally, total protein content and superoxide dismutase (SOD) activity were increased in response to AgNPs. Furthermore, bioactive compounds like gallic acid, tannic acid, coumarin, hesperidin, rutin, quercetin, and ferruginol were chromatographically separated and quantified using high-performance liquid chromatography (HPLC) with reference standards. These compounds were increased significantly in response to AgNPs treatments. We concluded that AgNPs could be a promising elicitor for improving the production of phytochemical compounds in medicinal plants. This work can serve asa good model for improving the production of bioactive compounds from medicinal plants in vitro. This molecular investigation should be done to understand better the metabolic mechanism leading to bioactive compound production scaling.

The distinctive characteristics of nanoparticles and their potential applications have been given considerable attention by scientists across different fields, particularly agriculture. However, there has been limited effort to assess the impact of copper nanoparticles (CuNPs) in modulating physiological and biochemical processes in response to salt-induced stress. This study aimed to synthesize CuNPs biologically using Solenostemma argel extract and determine their effects on morphophysiological parameters and antioxidant defense system of barley ( Hordeum vulgare ) under salt stress. The biosynthesized CuNPs were characterized by (UV–vis spectroscopy with Surface Plasmon Resonance at 320 nm, the crystalline nature of the formed NPs was verified via XRD, the FTIR recorded the presence of the functional groups, while TEM was confirmed the shape (spherical) and the sizes (9 to 18 nm) of biosynthesized CuNPs. Seeds of barley plants were grown in plastic pots and exposed to different levels of salt (0, 100 and 200 mM NaCl). Our findings revealed that the supplementation of CuNPs (0, 25 and 50 mg/L) to salinized barley significantly mitigate the negative impacts of salt stress and enhanced the plant growth-related parameters. High salinity level enhanced the oxidative damage by raising the concentrations of osmolytes (soluble protein, soluble sugar, and proline), malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ). In addition, increasing the activities of enzymatic antioxidants, total phenol, and flavonoids. Interestingly, exposing CuNPs on salt-stressed plants enhanced the plant-growth characteristics, photosynthetic pigments, and gas exchange parameters. Furthermore, CuNPs counteracted oxidative damage by lowering the accumulation of osmolytes, H 2 O 2 , MDA, total phenol, and flavonoids, while simultaneously enhancing the activities of antioxidant enzymes. In conclusion, the application of biosynthesized CuNPs presents a promising approach and sustainable strategy to enhance plant resistance to salinity stress, surpassing conventional methods in terms of environmental balance.

Among biological methods, green synthesis of the nanomaterials using plant extracts was shown to be an environmentally friendly, economical, and simple approach. In the current study, the biogenic synthesis of silver nanoparticles (AgNPs) was achieved using the leaf extract of Hibiscus tiliaceus, in order to prevent the contamination of the tissue culture media and induce callus growth. The nanostructures of the fabricated AgNPs were characterized using UV–visible spectroscopy, Fourier transform infra-red spectra (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), zeta size, and zeta potential techniques. Our results indicate that The UV–vis spectrum of AgNPs exhibited an absorption band at 415 nm. The FTIR analysis identified the functional groups which could involve in the reduction of silver ions to AgNPs, this was also confirmed by the (hkl) diffraction peaks in the XRD diffractogram. Moreover, the TEM analysis showed a spherical nanoparticle with a size ranging from 21 and 26 nm. Thereafter, the potential antibacterial and antifungal activity of the biogenic AgNPs was evaluated against Bacillus pumilus and Alternaria alternata which were isolated from the in vitro culture media and identified based on 16S rDNA and ITS rDNA sequences, respectively. The results showed that the AgNPs significantly inhibited the growth of Alternaria alternata and Bacillus pumilus at all applied concentrations (5, 10, 20 and 40 mg/L). Compared to the control more fungal radial growth reduction (42.59%,) and bacterial inhibition (98.12%) were registered in the plates containing high doses of AgNPs (40 mg/L). Using Rumex nervosus explants, the biosynthesized AgNPs were tested for their impact to promote callus growth. The obtained results showed a significant effect of AgNPs on callus fresh weight at all applied doses. Moreover, AgNPs treatments showed a polymorphism of 12.5% which was detected by RAPD markers. In summary, the results revealed that AgNPs (40 mg/L) can be effectively added to the in vitro culture media for reducing microbial contamination and improving callus growth while greatly maintaining its genetic stability.

Artemisia species are very important medicinal plants, particularly in the Middle East and in developing countries. Their products have been used in traditional and medicine contemporary for the treating of infectious ulcers, gangrenous ulcers, inflammations, and malaria. Artemisinin derived from Artemisia species has been used as a drug in many countries for malaria disease treatment. Hence, this study aimed to identify and evaluate the bioactive compounds of three species of Artemisia (Artemisia judaica, Artemisia monosperma, and Artemisia sieberi) growing in Saudi Arabia. Therefore, several analytical techniques, such as gas chromatography–mass spectrometry (GC-MS), UV-Visible spectrophotometry (UV-Vis), and high-performance liquid chromatography (HPLC), with reference standards, were used. The GC-MS analysis of the artemisia species revealed many bioactive constituents associated with plant secondary metabolites; some of these identified phytochemical components have biological activity. A. Judaica showed the highest number of bioactive compounds, followed by A. sieberi and A. monosperma. Further, the total phenol, total flavonoid, total tannin, terpenoids, and TCA were estimated. Furthermore, biomolecules such gallic acid, tannin acid, quercetin, and artemisinin in different artemisia species were quantified using HPLC with the reference standard. The amount of artemisinin in the leaf extract of these species (A. sieberi, A. Judaica, and A. monosperma) was found to be about 3.01, 2.5, and 1.9 mg/g DW, respectively. Moreover, the antioxidant activity of the samples was estimated. The obtained results have shown that these species possessed high antioxidant activity, and the scavenging of the DPPH radical and hydrogen peroxide were found to be raised with the increase in the plant extract concentration. This reflects the number of bioactive compounds in these species. The findings of this research support and justify the utilization of Artemisia species in folk medicine in the Middle East.

Drought stress is a significant abiotic stressor that has a negative impact on crop production and global food security systems. Drought stress was applied to eggplant seedlings with various field capacities (FC), 80% FC as control, 50% FC, 35% FC, and 20% FC. AgNPs were synthesised from green chemical methods, whereas different concentrations of AgNPs (0, 0.1, 0.2, 0.5 µmol) were applied exogenously on drought-stressed eggplants. Drought stress decreased the growth parameters (plant height, fresh mass, dry mass, leaf area), photosynthetic pigments (Chl a, Chl b, carotenoids), and protein content while increased the proline, hydrogen peroxide (H2O2), malondialdehyde (MDA) content, and activity of the antioxidant enzymes, i.e., superoxide dismutase (SOD) and catalase (CAT). AgNPs restricted proline accumulation and reduced H2O2, MDA content by upregulating the antioxidant enzymes. Overall, the current study’s findings indicated that AgNPs are an effective eco-friendly and low-cost application for plant growth under drought stress, with the potential to mitigate the impact of drought on plants.

Background Juniperus procera Hoechst . ex Endl. is a medicinal tree in Saudi Arabia, primarily in the Enemas region, but it is locally threatened due to die-back disease and difficulties regarding seed reproduction (seed dormancy and underdeveloped embryonic anatomy, and germination rate < 40%). Hence, the alternative methods for reproduction of Juniperus procera are really needed for conservation and getting mass propagation for pharmaceutical uses. Results In this manuscript, we articulated the successful in vitro shoot multiplication and callus induction of J. procera by using young seedling as explants and detected an important antibacterial and antitumor product. Explants were grown on different types of media with the supplement of different combinations of Plant Growth Regulators (PGRs) at different concentrations. The best media for shoot multiplication was Woody Plant Media (WPM) supplemented with PGRs (0.5 μM of IAA and 0.5 μM BAP or 0.5 μM IBA and 0.5 μM BAP). Whereas for callus induction and formation Woody Plant Media (WPM) with the addition of PGRs (0.5 μM 2,4-D and 0.5 μM BAP) was better than the Chu Basal Salt Mixture (N6), Gamborg’s B-5 Basal Medium (B5), and Murashige and Skoog media. The possibility of multiplication of J. procera in vitro creates significant advantages to overcome the difficulties of seeds dormancy for the reproduction of plants, conservation of trees, and getting mass propagation material for pharmaceutical studies. The shoot and callus extract of J. procera was detected using gas chromatography-mass spectrometry analysis and revealed more than 20 compounds related to secondary metabolites, which contained antibacterial and antitumor agents, such as ferruginol, Retinol, and Quinolone as well as confirmed by Direct Analysis in Real Time, Time of Flight Mass Spectrometry (DART-ToF-MS). Podophyllotoxin (PTOX) was detected in callus material by HPLC with sigma standard and confirmed by DART-ToF-MS and UV spectra. Conclusion We successfully conducted in vitro shoot multiplication and callus induction from J. procera seedlings using WPM and a different combination of PGRs and, detected an important antibacterial and antitumor product such as ferruginol and podophyllotoxin. According to our findings, J. procera has become a new natural source of novel bioactive compounds.

The implementation of nanotechnology in the field of plant tissue culture has demonstrated an interesting impact on in vitro plant growth and development. Furthermore, the plant tissue culture accompanying nanoparticles has been showed to be a reliable alternative for the biosynthesis of secondary metabolites. Herein, the effectiveness of zinc oxide nanoparticles (ZnONPs) on the growth of Delonix elata calli, as well as their phytochemical profiles, were investigated. Delonix elata seeds were collected and germinated, and then the plant species was determined based on the PCR product sequence of ITS1 and ITS4 primers. Afterward, the calli derived from Delonix elata seedlings were subjected to 0, 10, 20, 30, 40, and 50 mg/L of ZnONPs. The ZnONPs were biologically synthesized using the Ricinus communis aqueous leaf extract, which acts as a capping and reducing agent, and zinc nitrate solution. The nanostructures of the biogenic ZnONPs were confirmed using different techniques like UV–visible spectroscopy (UV), zeta potential measurement, Fourier transform infrared spectra (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). Adding 30 mg/L of ZnONPs to the MS media (containing 2.5 µM 2,4-D and 1 µM BAP) resulted in the highest callus fresh weight (5.65 g) compared to the control and other ZnONP treatments. Similarly, more phenolic accumulation (358.85 µg/g DW) and flavonoid (112.88 µg/g DW) contents were achieved at 30 mg/L. Furthermore, the high-performance liquid chromatography (HPLC) analysis showed significant increments in gallic acid, quercetin, hesperidin, and rutin in all treated ZnONP calli compared to the control. On the other hand, the gas chromatography and mass spectroscopy (GC-MS) analysis of the calli extracts revealed that nine phytochemical compounds were common among all extracts. Moreover, the most predominant compound found in calli treated with 20, 30, 40, and 50 mg/L of ZnONPs was bis(2-ethylhexyl) phthalate, with percentage areas of 27.33, 38.68, 22.66, and 17.98%, respectively. The predominant compounds in the control and in calli treated with 10 mg/L of ZnONPs were octadecanoic acid, 2-propenyl ester and heptanoic acid. In conclusion, in this study, green ZnONPs exerted beneficial effects on Delonix elata calli and improved their production of bioactive compounds, especially at a dose of 30 mg/L.

Pomegranate (Punica granatum L.) belongs to the Punicaceae family and is native to Central Asia; yet, it has a wide geographical distribution globally, reflecting its adaptation to different climatic conditions. Pomegranate is among the oldest and most significant cultivated crops, thriving extensively in tropical and subtropical climates. Besides its nutritional uses, pomegranate has been employed in traditional medicine for treating various diseases, including cancer prevention, antimicrobial activity, male infertility, ulcers, and diarrhea. The Bidah pomegranate cultivar is known for its unique sweet taste and high productivity yield. However, there is limited knowledge about its nutritional composition and medicinal value. Therefore, this study aimed to evaluate the functional potential of Bidah pomegranate in terms of its phytochemicals, antioxidant capacity, antibacterial, and anticancer activity. Different analytical techniques were used to investigate the chemical composition and antioxidant properties of Bidah pomegranate. Moreover, the biological activity of shoot and callus of Bidah pomegranate cultivar was assessed against Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and the colorectal cancer cell line (CaCo-2). Gas chromatography/mass spectrometry (GC/MS) analysis of the shoot and callus extracts revealed about 17 and 18 phytochemical compounds, respectively. Phenolic quantification showed that pomegranate materials contained high amounts of phenolic content. Additionally, Bidah pomegranate cultivar possesses high antioxidant activity with a low half-maximal inhibitory concentration (IC50) value. Furthermore, the pomegranate extract showed promising results with human pathogenic bacteria (E. coli and S. aureus), especially against S. aureus and the colorectal cancer cell line (CaCo-2). The findings of this study support the traditional use of pomegranate in folk medicine and highlight its potential for further exploration as a source of therapeutic agents.