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.

Green synthesis of zinc oxide nanoparticles (ZnO NPs) using plant extracts have recently attracted considerable attention due to their environmental protection benefits and their easy and low cost of fabrication. In the current study, ZnO NPS were synthesized using the aqueous extract of Ochradenus arabicus as a capping and reducing agent. The obtained ZnO NPs were firstly characterized using ultraviolet visible (UV-Vis) spectroscopy, Fourier transform infrared (FTIR), transmission electron microscope (TEM), X-ray diffraction (XRD), energy dispersive X-ray absorption (EDX), zeta potential, and zeta size. All these techniques confirmed the characteristic features of the biogenic synthesized ZnO NPs. Then, ZnO NPs were evaluated for their effects on morphological, biochemical, and physiological parameters of Salvia officinalis cultured in Murashige and Skoog medium containing 0, 75, 100, and 150 mM of NaCl. The results showed that ZnO NPs at a dose of 10 mg/L significantly increased the shoot number, shoot fresh weight, and shoot dry weight of Salvia officinalis subjected or not to the salt stress. For the shoot length, a slight increase of 4.3% was recorded in the plant treated by 150 mM NaCl+10 mg/L ZnO NPs compared to the plant treated only with 150 mM of NaCl. On the other hand, without NaCl, the application of both concentrations 10 mg/L and 30 mg/L of ZnO NPs significantly improved the total chlorophyll content by 30.3% and 21.8%, respectively. Under 150 mM of NaCl, the addition of 10 mg/L of ZnO NPs enhanced the total chlorophyll by 1.5 times, whilst a slight decrease of total chlorophyll was recorded in the plants treated by 150 mM NaCl + 30 mg/L ZnO NPs. Additionally, ZnO NPs significantly enhance the proline accumulation and the antioxidative enzyme activities of catalase (CAT), superoxide dismutase (SOD), and glutathione reductase (GR) in plants under salinity. Our findings revealed that green synthesized ZnO NPs, especially at a dose of 10 mg/L, play a crucial role in growth enhancement and salt stress mitigation. Hence, this biosynthesized ZnO NPs at a concentration of 10 mg/L can be considered as effective nanofertilizers for the plants grown in salty areas.

Zincoxide nanoparticles (ZnO NPs) are among the most produced and used nanomaterials worldwide, and in recent times these nanoparticles have also been incorporate in plant science and agricultural research. The present study was planned to synthesize ZnO NPs biologically using Ochradenus arabicus leaves and examine their effect on the morphology and physiology properties of Maerua oblongifolia cultured in vitro. ZnO NPs were characterized by UV–visible spectroscopy (UV–vis), X-ray diffractometer (XRD), Fourier transform infrared spectroscopy (FT-IR), and transmission electron microscopy, which demonstrated hexagonal shape nanoparticles of size ranging from 10 to 50 nm. Thus, the study uncovered an efficient, eco-friendly and simple technique for biosynthesis of multifunctional ZnO NPs using Ochradenus arabicus following growth of Maerua oblongifolia shoots in different concentrations of ZnO NPs (0, 1.25, 2.5, 5, 10, or 20 mg L⁻¹) in Murashige and Skoog medium. Remarkable increases in plant biomass, photosynthetic pigments, and total protein were recorded up to a concentration of 5 mg L⁻¹; at the same time, the results demonstrated a significant reduction in lipid peroxidation levels with respect to control. Interestingly, the levels of proline and the antioxidant enzyme catalase (CAT), superoxide dismutase (SOD), and glutathione reductase (GR) activities were increased significantly in response to all ZnO NP treatments. These findings indicate that bioengineered ZnO NPs play a major role in accumulation of biomass and stimulating the activities of antioxidant enzymes in plant tissues. Thus, green-synthesized ZnO NPs might be of agricultural and medicinal benefit owing to their impacts on plants in vitro.

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 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.