Differentiation of Mediterranean species of Juniperus from the Sabina section as a result of their migrations

The Sabina section is one of the three groups in the Juniperus genus and the most diverse. The variability of Mediterranean junipers from the Sabina section is related to their Tertiary and Pleistocene migrations and long-term isolations. Their contemporary taxonomic and geographic diversity was influenced by important events such as the migration of continents, the disappearance of Tethys, orogenic movements or the Messinian salinity crisis. The results of morphological measurements of seed cones, seeds and branchlets with leaves of 19 populations of Juniperus phoenicea complex, J. excelsa s.s., J. thurifera subsp. thurifera and subs. africana, J. foetidissima and J. sabina var. sabina and var. balkanensis were statistically compiled using univariate statistics and multivariate analysis. The most important characters differentiating the populations within the taxa were the thickness of the branchlet and the cone diameter, while between the taxa the ratio of cone diameter to the width of the seeds and the number of seeds per cone were used for speciation. J. phoenicea complex is distinguished from the other studied taxa by the greatest number of characters. J. foetidissima, J. sabina var. sabina and J. canariensis are characterized by the highest variability of morphological characters, while J. excelsa and J. sabina var. balkanensis – the lowest. The studies confirmed the ancient nature of the J. phoenicea complex in relation to other taxa from the Sabina section, as a result of an earlier detachment from the ancestor, and no loss of variability due to the effects of colonization and isolation in J. canariensis. In addition, the similarity of J. sabina and J. thurifera was demonstrated, which would confirm the descent from a common ancestor and similar migration routes from the center of Europe towards the Iberian Peninsula, as well as further differentiation of J. thurifera into subspecies caused by isolation due to the opening of the Strait of Gibraltar. The distinctiveness of J. foetidissima from all the other analyzed taxa was also confirmed, and some morphological similarity was shown, proving the original character of J. excelsa s.s. and its similarity to the J. phoenicea complex in this respect.

Preparation, Characterization, and Acetylcholinesterase Inhibitory Ability of the Inclusion Complex of β-Cyclodextrin–Cedar (Juniperus phoenicea) Essential Oil

The aim of the present study was the encapsulation of cedar (Juniperus phoenicea) essential oil (CEO) of Greek origin in β-cyclodextrin (β-CD) through the formation of inclusion complexes (ICs) using the co-precipitation method with different β-CD-to-CEO weight ratios (90:10, 85:15, 80:20, 70:30 (w/w)). The encapsulation of CEO in β-CD through host–guest interactions was confirmed by Nuclear Magnetic Resonance (NMR) spectroscopy, FT-IR spectroscopy, Differential Scanning Calorimetry (DSC) and Thermogravimetric Analysis (TGA). The obtained ICs exhibited nanoscale size (315.9 nm to 769.6 nm),Polydispersity Index from 0.326 to 0.604 and satisfactory stability in suspension (−37.0 mV to −17.0 mV). The process yield was satisfactory, ranging between 65% and 78%, while the inclusion efficiency ranged from 10% to 27%. The in vitro release study conducted for the IC with the optimal characteristics (β-CD:CEO 80:20 (w/w)) exhibited a sustained release profile, with an initial burst effect in the first 5 h. The release profile could be well expressed by the Higuchi equation: Q = 18.893 t1/2 + 9.5919, R2 = 0.8491. The cedar EO presented significant acetylcholinesterase inhibition (IC50 37 μg/mL), which was prolonged by its encapsulation into the β-CD cavity.

Phytochemical analysis, antioxidant and in vitro β-galactosidase inhibition activities of Juniperus phoenicea and Calicotome villosa methanolic extracts

Background Juniperus Phoenicea (JP) and Calicotome Villosa (CV) are used by Jordanian populations as herbal remedies in traditional medicine. Herein, the phytochemical contents of their methanolic extracts were analyzed and their antioxidant as well as in vitro anti- β-Galactosidase activities were evaluated; their effect on β-Galactosidase enzyme kinetics was evaluated and the thermodynamic of the enzyme was determined. Methods The antioxidant activity of JP and CV crude methanolic extracts was evaluated using 1,1-diphenyl,2-picrylhydrazyl (DPPH) free radical scavenging and ferric reducing antioxidant power (FRAP) assays; however, the effect of the plants’ crude extracts on β-Galactosidase activity and kinetics was evaluated in vitro. Moreover, total phenolic, flavonoids, and flavonols content in plants’ extracts were determined and expressed in Gallic acid equivalent (mg GAE/g dry extract) or rutin equivalent (mg RE/g dry extract). Results Phytochemical screening of the crude extracts of JP and CV leaves revealed the presence of phenols, alkaloids, flavonoids, terpenoids, anthraquinones, and glycosides. Flavonoids and flavonols contents were significantly higher in JP than in CV (p < 0.05). Furthermore, an analogous phenolic content was detected in both JP and CV methanolic extracts (103.6 vs 99.1 mg GAE/g extract). The ability of JP extract to scavenge DPPH radicals was significantly higher than that of CV extract with IC50 = 11.1 μg/ml and 15.6 μg/ml, respectively. However, their extracts revealed relatively similar antioxidant capacities in FRAP assay; their activity was concentration dependent. The JP extract inhibited β—galactosidase enzyme activity with a significant IC50 value compared to CV extract; they exhibited their inhibitory activities at IC50 values 65 µg/ml and 700 µg/ml, respectively. Rutin revealed anti-β-galactosidase activity at IC50 = 75 µg/ml. The mode of inhibition of β-galactosidase by JP, CV, and rutin was non-competitive, mixed, and competitive inhibition, respectively. Thermodynamic and enzyme inactivation kinetics revealed that β-galactosidase has a half-life time of 108 min at 55 °C, activation energy of 208.88 kJ mol−1 and the inactivation kinetics follows a first-order reaction with k-values 0.0023–0.0862 min−1 and positive entropy of inactivation (∆S°) values at various temperatures, indicating non-significant processes of aggregation. Conclusions The methanolic extracts of JP and CV possess anti-hyperglycemic and antioxidant activities with potential pharmaceutical applications.

Isolation, characterization, and antimicrobial activity of communic acid from Juniperus phoenicea

Objectives A mixture of Z and E communic acid is isolated for the first time from the cones of Juniperus phoenicea. Its biological activity was studied. Methods The plant material was extracted in a Soxhlet apparatus with n-hexane, the resulting extract was subjected to column chromatography (CC) on silica gel. The structure elucidation of the constituents of the isolated fraction was identified by comparison of its spectroscopic properties 1H and 13C NMR data with those reported in the literature. The antimicrobial assay of hexanic extract and isolated compounds was carried out by the disc diffusion and micro-dilution methods. Results A mixture of two diterpene acids isomers was isolated, with a high yield (68%). Their chemical structures were confirmed after comparing their spectral data with published reports. These natural products exhibited a significant antibacterial and antifungal activity against the tested strains. Indeed, for Bacillus cereus, Staphylococcus aureus, and Pseudomonas aeruginosa, the inhibition zone diameters (36–37 mm) was better than penicillin, novobiocin, and amoxicillin. For Candida albicans activity, it show that the mixture possess an activity similar to that of Metrazol. Against Escherichia coli, the inhibitory activity was found less than Amoxicillin. This is the first report of isolation of communic acid from J. phoenicea. Conclusions These results showed that the cones of J. phoenicea were an important source of communic acid, and its hexanic extract had the greatest potential antibacterial activity against both Gram-negative and Gram-positive bacteria and C. albicans.

Evaluation of antioxidants activity of some tree barks grown in Libya (Al Jabal Al Akhdar) Pinus halepensis Mill, Pistacia lentiscus L, Juniperus phoenicea L

The aim of the present study investigated anti-oxidant activity of various methanol-water extracts from tree barks grown in Libya (Al Jabal AL Akhdar) by four anti-oxidant assays FRAP. DPPH. H2O2 and Metal chelating activity. Additionally, these antioxidant activities were compared with BHA, BHT as reference antioxidants. Tree bark has a large, diverse class of compounds, many with antioxidant properties. This study showed FRAP inhibitory activity of the Pinus halepensis bark extracts 56.8%, while Juniperus phoenicea L 57.6%, and Pistacia lentiscus L bark extracts was 69.2% The DPPH radical scavenging in the bark extracts exerted an inhibition of 66.8 %, 62.3% ,74.6% for Pinus halepensis, Juniperus phoenicea and Pistacia lentiscus L respectively. While H2O2 activity shows variation, ranging from 67. 8 to 81. 3%. The Metal chelating activity of the barks extract was 59% in Pinus halepensis and 57% in Juniperus phoenicea L bark extracts furthermore Pistacia lentiscus L was 67.8% in addition the Metal chelating activity of BHT and BHA 92%, 94% respectively. The high antioxidant activity of bark was founded in Pistacia lentiscus L bark extracts.

Chemical Composition and Antimicrobial Activity of Essential oil from Scales of Moroccan Juniperus phoenicea

Abstract-The essential oil of Juniperus phoenicea was obtained by hydrodistillation method using a Clevenger-type apparatus with a yield of 1.9 % and was analyzed by gas chromatography coupled to a mass spectrometer (GC-MS). Twelve volatile compounds were identified representing 99,85% of the total oil composition, while the α-pinene (78,31%), β-Myrcene (11,92%) and limonene (3,96%) were the major compounds. This essential oil was evaluated as an antibacterial and antifungal agent. The result showed that the oil presents a high biological activity as an antibacterial agent against the three tested strains Escherichia coli, Staphylococcus aureus and Pseudomonas aeroginos. It's also active as an antifungal agent against the Candida albicans with a zone inhibition of 28 mm. Keywords: Medicinal plant, Juniperus, phoenicea, Essential oil, Chemical composition, Antimicrobial activity.

In situ approaches show the limitation of the spoilage potential of Juniperus phoenicea L. essential oil against cold-tolerant Pseudomonas fluorescens KM24

The present study aimed to elucidate the effect of subinhibitory concentrations (sub-MICs) of juniper essential oil (EO), α-pinene, and sabinene on the quorum-sensing (QS)–mediated proteolytic and lipolytic properties of Pseudomonas fluorescens KM24. These activities were verified under in situ conditions, in which sub-MICs of the agents altered the morphology of KM24 cells. RNA-Seq studies revealed key coding sequences (CDSs)/genes related to QS and the proteolytic/lipolytic activities of pseudomonads. In this work, all the examined agents decreased autoinducer synthesis and influenced the mRNA expression of the encoding acyltransferase genes lptA, lptD, and plsB. The highest reduction on the 3rd and 5th days of cultivation was observed for the genes lptD (−5.5 and −5.61, respectively) and lptA (−3.5 and −4.0, respectively) following treatment with EO. Inhibition of the lptA, lptD, and plsB genes by singular constituents of EO was on average, from −0.4 to −0.7. At 5 days of cultivation the profile of AHLs of the reference P. fluorescens KM24 strain consisted of 3-oxo-C14-HSL, 3-oxo-C6-HSL, C4-HSL, and N-[(RS)-3-hydroxybutyryl]-HSL, the concentrations of which were 0.570, 0.018, 3.744, and 0.554 μg ml−1, respectively. Independent of the incubation time, EO, α-pinene, and sabinene also suppressed the protease genes prlC (−1.5, −0.5, and −0.5, respectively) and ctpB (−1.5, −0.7, and −0.4, respectively). Lipolysis and transcription of the lipA/lipB genes were downregulated by the agents on average from −0.3 to −0.6. α-Pinene- and sabinene-rich juniper EO acts as an anti-quorum-sensing agent and can repress the spoilage phenotype of pseudomonads. Key points: Juniper EO, α-pinene, sabinene exhibited anti-QS potential toward KM24. RNA-Seq revealed key CDSs/genes related to QS/proteolytic/lipolytic activities of KM24. Agents at sub-MIC levels influenced the mRNA expression of QS/lipase/protease genes.