Pomegranate Leaf Research Articles

Physiological and metabolomic analysis of Punica granatum (L.) under drought stress

Punica granatum has a noticeable adaptation to drought stress. The levels of the green leaf volatile trans-2-hexenal increased in response to drought stress suggesting a possible role of this compound in drought stress response in pomegranate. Punica granatum (L.) is a highly valued fruit crop for its health-promoting effects and it is mainly cultivated in semi-arid areas. Thus, understanding the response mechanisms to drought stress is of great importance. In the present research, a metabolomics analysis was performed to evaluate the effects of drought stress on volatile organic compounds extracted from the leaves of pomegranate plants grown under water shortage conditions. The time course experiment (7 days of water deprivation and 24-h recovery) consisted of three treatments (control, drought stress, and rehydration of drought-stressed plants). Plant weights were recorded and control plants were irrigated daily at pot capacity to provide the lost water. Fraction of transpirable soil water has been evaluated as indicator of soil water availability in stressed plants. The levels of proline, hydrogen peroxide and lipid peroxidation as well as of the photosynthetic parameters such as photosynthesis rate (A), stomatal conductance (g s), photosynthetic efficiency of photosystem II, and photochemical quenching were monitored after the imposition of drought stress and recovery as markers of plant stress. Constitutive carbon volatile components were analyzed in the leaf of control and drought-stressed leaves using Head Space Solid Phase Micro Extraction sampling coupled with Gas Chromatography Mass Spectrometry. A total of 12 volatile compounds were found in pomegranate leaf profiles, mainly aldehydes, alcohols, and organic acids. Among them, the trans-2-hexenal showed a significant increase in water-stressed and recovered leaves respect to the well-watered ones. These data evidence a possible role of the oxylipin pathway in the response to water stress in pomegranate plants.

Characterization of green zero-valent iron nanoparticles produced with tree leaf extracts

In the last decades nanotechnology has become increasingly important because it offers indisputable advantages to almost every area of expertise, including environmental remediation. In this area the synthesis of highly reactive nanomaterials (e.g. zero-valent iron nanoparticles, nZVI) is gaining the attention of the scientific community, service providers and other stakeholders.The synthesis of nZVI by the recently developed green bottom-up method is extremely promising. However, the lack of information about the characteristics of the synthetized particles hinders a wider and more extensive application.This work aims to evaluate the characteristics of nZVI synthesized through the green method using leaves from different trees. Considering the requirements of a product for environmental remediation the following characteristics were studied: size, shape, reactivity and agglomeration tendency.The mulberry and pomegranate leaf extracts produced the smallest nZVIs (5–10nm), the peach, pear and vine leaf extracts produced the most reactive nZVIs while the ones produced with passion fruit, medlar and cherry extracts did not settle at high nZVI concentrations (931 and 266ppm). Considering all tests, the nZVIs obtained from medlar and vine leaf extracts are the ones that could present better performances in the environmental remediation.The information gathered in this paper will be useful to choose the most appropriate leaf extracts and operational conditions for the application of the green nZVIs in environmental remediation.

Assessment of Antimicrobial Activity of Pomegranate Leaf Extract against Bacterial Isolates of Otitis Externa and Diarrhea of Dogs

Bacterial pathogens were isolated from cases of otitis externa and diarrhoea in dogs at Durg and Raipur districts of Chhattisgarh. All isolates were further subjected for antibiogram profile using commonly used antibiotics. Total 105 bacterial isolates were obtained which included Staphylococcus sp. and E. coli as predominant pathogens followed by Proteus, Klebsiella, Pseudomonas, Micrococcus and Salmonella. In cases of otitis externa, ciprofloxacin was the most effective antibiotic (67.27%) followed by Gentamicin (61.81%), Cephalaxin (40%), Chloramphenicol (32.72%), Amoxycillin (21.81%), Ampicillin (20%), Doxycycline (16.36%), Penicillin (12.72%) and Erythromycin (3.63%). However, Chloromphenical (72%) was reported as the most effective drug against bacterial isolates of diarrhoea followed by Ciprofloxacin (70%), Gentamicin (52%), Cephalaxin (40%), Doxycyline (30%), Ampicillin (14%), Amoxycillin (6%) and Erythromycin (2%). Cold extract of pomegranate leaves was then assessed for antibacterial activity against all isolates by disc diffusion and tube dilution methods. The zones of inhibition of pomegranate leaves extract (PLE) against Staphylococcus aureus, Pseudomonas aeruginosa, Proteus, Klebsiella, E. coli and Salmonella were 18.8 ± 0.050, 5±0.70, 7.8±0.73, 16.2±0.86, 15.6±0.92 and 15.2±0.86 mm respectively. The zone of inhibition with Staphylococcus was significantly higher (P<0.05) than other bacteria. The minimum bactericidal concentration against Staphylococcus aureus, Klebsiella, E. coli and Salmonella were 33.33±8.34, 66.66±16.68, 66.66±16.68 and 33.33±8.34 mg/ml respectively. Staphylococcus aureus showed maximum sensitivity to pomegranate leaf extract amongst all the organisms studied, followed by Klebsiella, E. coli and Salmonella. Although, Proteus and Pseudomonas were refractory to PLE, but moderate sensitivity was noticed even against few multidrug resistant isolates.

PRELIMINARY AND PHARMACOGNOSTIC INVESTIGATION ON LEAVES OF PUNICA GRANATUM

Among the medicinal plants Punica granatum commonly known as pomegranate is one of the important ethno medicinal plants used in traditional and folklore therapies. For the preparation of aqueous extract decoction method, methanolic and hexane extract soxhlet distillation method s were used. The methanolic crude extract show higher degree of inhibitory activity against Escheri chia coli , Staphylococcus aureus and Salmonella typhi . Phyto chemical analysis conducted on whole leaf extracts of Punica granatum revealed the presence of various chemical constituents having medicinal values. Through proximate analysis identification of plant material and to ascertain it’s quality and purity. In the present study morphology, macroscopic, microscopic, physico chemical, proximate, phytochemical screening of po wdered crude drug were carried out as referential information for identification and characterization of Punica granatum leaf and it’s extracts.

Facile synthesis of size-tunable gold nanoparticles by pomegranate (Punica granatum) leaf extract: Applications in arsenate sensing

When pomegranate leaf extracts were incubated with chloroauric acid (HAuCl4), gold nanoparticles (AuNPs) were synthesized. These were characterized by a variety of techniques. With an increasing content of the leaf extract, a gradual decrease in size and an increase in monodispersity were observed. Transmission electron microscope (TEM) images showed that the phyto-fabricated AuNPs were surrounded by an amorphous layer. Gallic acid in the extract mediated the reduction and a natural decapeptide capped the nanostructures. Blocking of thiol groups in the decapeptide cysteine residues caused the nanoparticles to aggregate. On interaction with arsenate (V) ions, the UV–vis spectra of the nanoparticles showed a decrease in intensity and a red-shift. Energy dispersive spectra confirmed the presence of arsenate associated with the AuNPs. Thus, by using these AuNPs, a method for sensing the toxic arsenate ions could be developed.

Anti-oxidant Activity and Whitening Activity of Punica granatum Leaf Extracts

Anti-oxidant Activity and Whitening Activity of Punica granatum Leaf Extracts

In vitro and in vivo evaluation of antioxidant and antigenotoxic potential of Punica granatum leaf extract

Context: Several studies have reported the antioxidant activity and potential therapeutic properties of Punica granatum L. (Lythraceae) fruit. Medicinal properties have also been attributed to other parts of P. granatum tree, which are rich in bioactive phytochemicals.Objective: To explore the phytochemical characteristics, in vitro and in vivo antioxidant and in vivo antigenotoxic potential of P. granatum leaf extract (PLE).Materials and methods: The in vitro antioxidant potential of PLE was assessed by DPPH (1,1-diphenyl-2-picrylhydrazyl), ferric reducing antioxidant power (FRAP). Inhibition of lipid peroxidation (LPO) and the total phenolic content of the samples were also determined. Thirty-six male Swiss albino mice were divided into six groups (six animals each). Group 1 (control) and group 2 mice received vehicle and genotoxin alone, respectively. Groups 3, 4 and 5 were pretreated with PLE (400, 600 and 800 mg/kg body weight, respectively) prior to the administration of genotoxin. Group 6 received highest test dose of PLE. DNA damage in the bone marrow cells, hepatic LPO and antioxidants were recorded.Results: Phytochemical analysis of PLE showed the presence of flavonoids, phenols, phytosterols, tannins and carbohydrates. Aqueous PLE demonstrated free radical scavenging activity, reducing power and inhibition of LPO with the EC50 values of 10.25, 59.88 and 20.05, respectively. A significant protective effect was observed against cyclophosphamide induced DNA damage and inhibition of hepatic LPO with concomitant increase in reduced glutathione (GSH) glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) in mice pretreated with PLE.Discussion and conclusion: PLE demonstrated a significant antioxidant and antigenotoxic potential and hence can be a potential natural source in health and medicine.

Phyto-inspired Silica Nanowires: Characterization and Application in Lipase Immobilization

Silica nanostructures were phyto-fabricated on different surfaces by using pomegranate (Punica granatum) leaf extracts. On zinc films, nanowires were obtained. On other surfaces such as silica, alumina, zinc oxide, and glass, spherical aggregates, cubic assemblies, microflakes, and acicular structures, respectively, were observed. The nanowires developed on Zn surfaces were characterized by scanning electron microscope-energy dispersive spectrometer (SEM-EDS), X-ray diffraction (XRD), photoluminescence, and Fourier transform infrared (FTIR) spectroscopic analysis. XRD profiles displayed peaks at 2.4, 4.9, and 12.1° indicating the presence of silica nanostructures. When excited at 340 nm, the reaction mixtures displayed a characteristic blue luminescence at 404 nm. FTIR spectra showed the existence of Si-OH and Si-O-Si bonds. The nanowires were functionalized with amine groups and used for the covalent immobilization of Candida rugosa lipase. The immobilized enzyme displayed better pH and temperature stability and retained 80% activity after 20 cycles. This paper highlights a novel route for the phyto-mediated growth of silica nanowires on Zn surfaces, their characterization and effective use as a matrix for enzyme immobilization.

High potential of agro-industrial by-products of pomegranate ( Punica granatum L.) as the powerful antifungal and antioxidant substances

Microorganisms such as fungi are one of the most important factors that cause oxidative processes during postharvest stage and consequently deterioration of agriculture products would not be unexpected. On the other hand, high antioxidant properties of industrial by-products of pomegranate propose them as powerful antioxidant and antifungal substances. So to investigate the antioxidant and antifungal properties of pomegranate, two independent factorial experiments based on randomized design with 5 replications were conducted. In the first experiment the effect of 3 different parts of pomegranate (peel, seed and leaf) and 2 different kinds of extracts (aqueous and methanolic) with 4 concentrations (0, 500, 1000 and 1500 ppm) were investigated on 3 postharvest fungi ( Penicillium italicum, Rhizopus stolonifer and Botrytis cinerea). In the second experiment antioxidant capacity and phenolic content were measured for two different extracts from different parts. Based on the results the methanolic extract showed the highest inhibitory effects on the mycelia growth (IMG) and spore germination (ISG) with 49.82 and 41.25% respectively. On the other hand, peel and seed extracts had more inhibitory effect (IMG and ISG) than leaf extract. The phenolic content of peel extract were also measured 2.8 fold higher than pomegranate leaf extract and antioxidant capacity of peel, seed and leaf extracts of pomegranate were 55.3%, 35.7% and 16.4% respectively. Therefore, it seems that the high percentage of phenolic content in the peel and seed of pomegranate could cause the high antifungal and antioxidant activity of their extracts.

Effect of Extraction Techniques and Solvents on Antioxidant Activity of Pomegranate (Punica granatum L.) Leaf and Stem

The effect of commonly used techniques and solvents in the antioxidant activities of Pomegranate leaves and stems were studied. The extraction techniques compared were successive, individual (cold percolation), and decoction methods in both parts using solvents viz. petroleum ether, toluene, ethyl acetate, acetone, and water. The antioxidant activity of all the different solvent extracts of leaves and stem was evaluated using antioxidant assays like 2,2-diphenyl-1-picryl hydrazyl free radical scavenging assay, superoxide anion radical scavenging assay and ferric-reducing antioxidant power. Total phenol and flavonoid content was also measured. Successive extraction was a better technique to extract the antioxidants from pomegranate than other techniques evaluated in the present study. For the aqueous extraction, decoction method was found to be the best method to extract the antioxidants and it is the most convenient, exhaustive, and time-saving extraction technique for both parts of pomegranate. The results showed that the extracting solvent significantly altered the antioxidant property estimations of pomegranate leaf and stem. High correlations between phenolic compositions and antioxidant activities of pomegranate extracts were observed. High levels of antioxidant activities were detected in pomegranate leaf as compared to stem indicating that the leaf may serve as an excellent source of natural antioxidants.

Pomegranate leaves and mulberry fruit as natural sensitizers for dye-sensitized solar cells

This study employs chlorophyll extract from pomegranate leaf and anthocyanin extract from mulberry fruit as the natural dyes for a dye-sensitized solar cell (DSSC). A self-developed nanofluid synthesis system is employed to prepare TiO 2 nanofluid with an average particle size of 25 nm. Electrophoresis deposition was performed to deposit TiO 2 nanoparticles on the indium tin oxide (ITO) conductive glass, forming a TiO 2 thin film with the thickness of 11 μm. Furthermore, this TiO 2 thin film was sintered at 450 °C to enhance the thin film compactness. Sputtering was used to prepare counter electrode by depositing Pt thin film on FTO glass at a thickness of 20 nm. The electrodes, electrolyte ( I 3 - ), and dyes were assembled into a cell module and illuminated by a light source simulating AM 1.5 with a light strength of 100 mW/cm 2 to measure the photoelectric conversion efficiency of the prepared DSSCs. According to experimental results, the conversion efficiency of the DSSCs prepared by chlorophyll dyes from pomegranate leaf extract is 0.597%, with open-circuit voltage ( V OC ) of 0.56 V, short-circuit current density ( J SC ) of 2.05 mA/cm 2, and fill factor (FF) of 0.52. The conversion efficiency of the DSSCs prepared by anthocyanin dyes from mulberry extract is 0.548%, with V OC of 0.555 V and J SC of 1.89 mA/cm 2 and FF of 0.53. The conversion efficiency is 0.722% for chlorophyll and anthocyanin as the dye mixture, with V OC of 0.53 V, J SC of 2.8 mA/cm 2, and FF of 0.49.

Genotypic characterization of Bacillus subtilis strains colonizing pomegranate phyllosphere and their antagonistic activities against Cercospora beticola

The community composition of epiphytic heterotrophic Bacillus subtilis on leaves of pomegranate was investigated. The bacteria were isolated on nutrient agar and systematically identified with morphological characters. Fifteen strains were characterized for their antagonistic potential against Cercospora beticola (causal agent for pomegranate leaf spot). All tested strains showed a significant inhibitory effect against the pathogen. Randomly amplified polymorphic DNA (RAPD) fingerprinting technique was applied to study the genetic diversity among the tested strains. This study indicated that there is a significant biochemical and genomic diversity among the B. subtilis strains and this diversity is not highly correlated with habitats and the antagonistic effect of the studied strains.

Transport behavior of ellagic acid of pomegranate leaf tannins and its correlation with total cholesterol alteration in HepG2 cells

The aim of this study was to investigate whether ellagic acid in pomegranate leaf tannins could be transported into HepG2 cells and its transport behavior. High-performance liquid chromatography coupled with a 996 photodiode array detector at 254 nm was applied. The mobile phase was an acetonitrile-water solution (containing 0.1% triethylamine, pH 3.0; 16:64, v/v, for determining ellagic acid in cells). The flow rate was 0.8 mL/min. Cells were incubated with pomegranate leaf tannins with 100 and 50 microg/mL (containing 1.71 and 0.85 microg/mL of ellagic acid, respectively) for a specific time, then lysed and sonicated in methanol to extract intracellular ellagic acid. A 10 microL aliquot of sample was injected into the HPLC system to determine ellagic acid concentration. The results showed that ellagic acid in pomegranate leaf tannins could be transported into the cells, which was in correlation with total cholesterol alteration in the cells. This is the first time that the transport behavior of ellagic acid through HepG2 cells in vitro has been comprehensively demonstrated.

Effects of Season, Variety, and Processing Method on Ellagic Acid Content in Pomegranate Leaves

Ellagic acid (EA) has aroused great interest worldwide owing to its antioxidant, anti-inflammatory, and anticarcinogenetic properties. The EA content in pomegranate leaf was measured in this study using high performance liquid chromatography to investigate the effects of season, variety, and processing method on the EA level. The results show that the EA content in 11 varieties of pomegranate from the Zaozhuang region in China range from 1.30 mg · g −1 to 6.46 mg · g −1 of dry weight in five consecutive seasons from June to October. An analysis of variance (ANOVA) shows that the EA content is significantly dependent on the season ( p<0.05). The EA content increases significantly during the growing season to the highest level in September and October. The effect of the leaf variety on the EA content is less significant than the season. The processing methods have different effects on the EA content. Soaking for 24 hours slightly increases the EA content ( p<0.05). Heating at 80°C or 100°C for 1 h after soaking has little influence on the EA content, while slow-fired cooking at high temperature significantly elevates the EA content ( p<0.05). To improve quality and stability, several parameters such as leaf collection time, slow-fired cooking, and cooking time should be strictly controlled during the processing of pomegranate leaf tea and its extract.

Evidence of anti-obesity effects of the pomegranate leaf extract in high-fat diet induced obese mice

To investigate the anti-obesity effects of the pomegranate leaf extract (PLE) in a mouse model of high-fat diet induced obesity and hyperlipidemia. For the anti-obesity experiment, male and female ICR mice were fed with a high-fat diet to induce obesity. When the weight of the high-fat diet group was 20% higher than the normal diet group, the animals were treated with 400 or 800 mg/kg/day of PLE for 5 weeks. Body weight and daily food intake were measured regularly during the experimental period. The various adipose pads were weighed and serum total cholesterol (TC), triglyceride (TG), glucose and high-density lipoprotein cholesterol (HDL-C) were measured after 5 weeks, treatment with PLE. In the fat absorption experiment, both the normal and obese mice were given 0.5 ml lipid emulsion and PLE at a dose of 800 mg/kg at the same time. Serial serum TG levels were measured at times 1, 2, 3, 4 and 6 h after the treatment. TGs in fecal excretions were measured after the mice were orally given a lipid emulsion. Effects of PLE and its isolated compounds (ellagic acid and tannic acid) on pancreatic lipase activity were examined in vitro. The PLE-treated groups showed a significant decrease in body weight, energy intake and various adipose pad weight percents and serum, TC, TG, glucose levels and TC/HDL-C ratio after 5 weeks treatment. Furthermore, PLE significantly attenuated the raising of the serum TG level and inhibited the intestinal fat absorption in mice given a fat emulsion orally. PLE showed a significant difference in decreasing the appetite of obese mice fed a high-fat diet, but showed no effect in mice fed a normal diet. PLE can inhibit the development of obesity and hyperlipidemia in high-fat diet induced obese mice. The effects appear to be partly mediated by inhibiting the pancreatic lipase activity and suppressing energy intake. PLE may be a novel appetite suppressant that only affects obesity owing to a high-fat diet.

Pharmacokinetic study of ellagic acid in rat after oral administration of pomegranate leaf extract

Quantification of ellagic acid, the principal bioactive component of pomegranate leaf extract, in rats plasma following oral administration of pomegranate leaf extract was achieved by using a high-performance liquid chromatographic method. The calibration curve for ellagic acid was linear ( r 2=0.9998) ver the concentration range 0.026–1.3 μg/ml. The intra- and inter-day assays of ellagic acid from rat plasma were less than 6.52% at concentration range from 26 to 1300 ng/ml and good overall recoveries (94.5–102.4%) were found on same concentrations. The concentration–time profile was fitted with an open two-compartment system with lag time and its max concentration of ellagic acid in plasma was 213 ng/ml only 0.55 h after oral administration extract 0.8 g/kg. The pharmacokinetic profile indicates that ellagic acid has poor absorption and rapid elimination after oral administration pomegranate leaf extract, and part of it was absorbed from stomach.

Adventitious Shoot and Plantlet Formation from Cultured Pomegranate Leaf Explants

Abstract Callus from leaf explants of two pomegranate (Punica granatum L.) clones produced adventitious shoots after 2 months of in vitro culture. Leaf segment explants initially were cultured on a modified Murashige and Skoog (MS) basal medium supplemented with a range of 0.5-10 μm BA and 0.05-5 (μm NAA. Shoot elongation was stimulated most efficiently when the initial calli were transferred from the shoot induction medium to half-strength MS supplemented with 2 μm BA. Elongated shoots rooted easily on a medium of one-half MS and 0.1 μm NAA. Whole plants were recovered after transfer to vermiculite in a greenhouse. Chemical names used: N-(phenylmethyl)-1H-purin-6-amine (BA), 1-naphthaleneacetic acid (NAA).