Chemical Constituents and Anticancer Activities of the Extracts from Phlomis × commixta Rech. f. (P. cretica × P. lanata).

The present work is the first report on the ingredients of the P. × commixta hybrid, a plant of the genus Phlomis. So far, thirty substances have been isolated by various chromatographic techniques and identified by spectroscopic methods, such as UV/Vis, NMR, GC-MS and LC-MS. The compounds are classified as flavonoids: naringenin, eriodyctiol, eriodyctiol-7-O-ß-D-glucoside, luteolin, luteolin-7-O-ß-D-glucoside, apigenin, apigenin-7-O-ß-D-glucoside, diosmetin-7-O-ß-D-glucoside, quercetin, hesperetin and quercetin-3-O-ß-D-glucoside; phenylpropanoids: martynoside, verbascoside, forsythoside B, echinacoside and allysonoside; chromene: 5,7-dihydroxychromone; phenolic acids: caffeic acid, p-hydroxybenzoic acid, chlorogenic acid, chlorogenic acid methyl ester, gallic acid, p-coumaric acid and vanillic acid; aliphatic hydrocarbon: docos-1-ene; steroids: brassicasterol and stigmasterol; a glucoside of allylic alcohol, 3-O-ß-D-apiofuranosyl-(1→6)-O-ß-D-glucopyranosyl-oct-1-ene-3-ol, was fully characterized as a natural product for the first time. Two tyrosol esters were also isolated: tyrosol lignocerate and tyrosol methyl ether palmitate, the latter one being isolated as a natural product for the first time. Moreover, the biological activities of the extracts from the different polarities of the roots, leaves and flowers were estimated for their cytotoxic potency. All root extracts tested showed a high cytotoxic activity against the Hep2c and RD cell lines.

Phytochemicals, Essential Oils Composition and Antioxidant Activity of Astragalus spp., Phlomis olivieri and Daphne mucronata in Habitats of Central Iran.

This study evaluated several secondary metabolites, essential oils (EOs) compositions, and antioxidant activity in four medicinal plants that originated in Isfahan rangelands. The species were Astragalus verus, Astragalus adscendens, Daphne mucronata, and Phlomis olivieri. Thirty-two genotypes of these species were evaluated for different biochemical traits. Based on the evaluation of EOs compounds, GC/MS analysis revealed the total number of identified compounds. These compounds were 25, 22, 12, and 22 for A. adscendens, A. verus, D. mucronata, and P. olivieri, respectively. The dominant compounds were phthalate (59.88 %) in A. adscendens, phytol (38.02 %) in A. verus, hexanoic acid (32.05 %) in D. mucronata and ß-cubebene (30.94 %) in P. olivieri. Phytochemical analysis showed that D. mucronata, A. adscendens, and P. olivieri had the highest total phenolics content (TPC) (18.24 mg gallic acid equivalent/g dry weight), total flavonoids content (5.57 mg QE/g DW), and total anthocyanins content (0.23 mg/g DW), respectively. The highest total chlorophyll (0.27 mg/g DW), total carotenoids (0.03 mg/g DW), and antioxidant activity (71.36 %) were observed in A. adscendens, A. adscendens and A. verus, respectively. Among all genotypes, the highest TPC (20.1 mg GAE/g DW) was observed in genotype 5 of D. mucronata. This study provided new information on the chemical compounds within the distribution range of these ecologically dominant rangeland species in Isfahan province, Iran. The data revealed that superior genotypes from these species are rich in natural antioxidants and bioactive compounds. Thus, they can be used in ethno pharmacological fields, food, and industrial applications.

In Vitro/In Vivo Hepatoprotective and Antioxidant Effects of Defatted Extract and a Phenolic Fraction Obtained from Phlomis Tuberosa.

An in vitro/in vivo hepatotoxicity and hepatoprotection evaluation of a defatted extract and a phenolic fraction from Phlomis tuberosa, administered alone and in a carbon tetrachloride (CCl4)-induced metabolic bioactivation model, was performed. The extract and the phenolic fraction were analysed by high performance liquid chromatography (HPLC) to determine the total flavonoid content, to identify flavonoids and to quantify verbascoside. In addition, total polyphenolics in the samples were expressed as gallic acid equivalents. Applied alone, the extract and the fraction (5, 10 and 50 µg/mL) did not show a statistically significant hepatotoxic effect on isolated rat hepatocytes in vitro. In a CCl4-induced hepatotoxicity model, the samples exhibited a concentration-dependent, statistically significant hepatoprotective effect, which was most pronounced at 50 µg/mL for both. The phenolic fraction exhibited a more pronounced hepatoprotective effect compared to the extract. Data from the in vitro study on the effects of the extract were also confirmed in the in vivo experiment conducted in a CCl4-induced hepatotoxicity model in rats. A histopathological study showed that the animals treated with CCl4 and the extract had an unaltered histoarchitecture of the liver. The effects of the extract were the same as those of silymarin.

Phytochemical analysis and antibacterial/antifungal activity of the essential oil of Phlomis olivieri Benth in Iran.

Phlomis olivieri Benth. is an aromatic plant endemic to Iran belonging to the Lamiaceae family. It is used to treat pain, stomach ache and common cold in Iranian traditional medicine. P. olivieri also has valuable biological properties including antioxidant, antimicrobial and analgesic ones. This was the first study designed to assess the quality, quantity and antimicrobial activity of Phlomis olivieri Benth. essential oil (POEO). Samples were randomly collected from flowering twigs of this species in three locations between Azeran and Kamoo in Kashan, Iran at peak flowering in June 2019. Water distillation extraction was used to obtain the POEO the quantity of which was calculated by weight. Gas chromatography coupled to mass spectrometry (GC/MS) was also used for POEO qualitative analysis, which revealed its chemical compounds and their percentages. Antimicrobial activity of POEO was also determined using the agar well diffusion method. Minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentration (MBC/MFC) were also measured using the broth microdilution method. The results of the quantitative and qualitative analysis showed that the POEO yield was ~ 0.2292% and its main chemical compounds included the sesquiterpenes germacrene D (26.43%), ß-caryophyllene (20.72%), elixene (6.58%), ß-trans-farnesene (6.17%), ß-Cyclogermacrane (5.04%), germacrene B (4.73%), α-humulene (4.22%), and monoterpene α-pinene (3.22%). The agar diffusion method demonstrated the highest antimicrobial activity of POEO (MIC ~ 14.50 mm) was against the Gram-positive species Streptococcus pyogenes. The POEO also showed the strongest inhibitory and lethal activity against the gram-negative bacterial species Pseudomonas aeruginosa (MIC < 62.50 µg/mL) and S. paratyphi-A (MIC < 62.50 µg/mL and MBC = 125 µg/mL), and fungal species Candida albicans (MIC and MBC = 250 µg/mL) as compared to control-positive antibiotics. Therefore, POEO is a valuable natural alternative rich in sesquiterpenes with strong antimicrobial and antifungal activities against some fungal and bacterial strains. It can also be used in the pharmaceutical, food and cosmetic industries.

Anti-asthmatic effects of Phlomis umbrosa Turczaninow using ovalbumin induced asthma murine model and network pharmacology analysis.

BACKGROUND: Phlomis umbrosa Turczaninow has been used as a tradition herbal medicine for treating various inflammatory diseases. PURPOSE: In present study, we explored the effects of P. umbrosa on asthma induced by ovalbumin (OVA) and elucidated the mechanism via in vivo verification and network pharmacology prediction. METHODS: The animals were intraperitoneally injected OVA on day 1 and 14, followed by OVA inhalation on days 21, 22, and 23. The animals were daily treated P. umbrosa extract (PUE, 20 and 40 mg/kg) by oral gavage from day 18 to day 23. RESULTS: PUE significantly decreased airway hyperresponsiveness, eosinophilia, and the production of inflammatory cytokines and OVA specific immunoglobulin E in animals with asthma, along with a reduction in airway inflammation and mucus secretion in lung tissue. In network analysis, antiasthmatic effects of PUE were closely related with suppression of mitogen-activated protein kinases and matrix metalloproteinases (MMPs). Consistent with the results from network analysis, PUE suppressed the phosphorylation of ERK and p65, which was accompanied by a decline in MMP-9 expression. CONCLUSION: Administration of PUE effectively reduced allergic responses in asthmatic mice, which was associated with the suppressed phosphorylation of ERK and p65, and expression of MMP-9. These results indicate that PUE has therapeutic potential to treat allergic asthma.

Isolation of microsatellite markers for the endemic Phlomis lychnitis (Lamiaceae).

BACKGROUND: Phlomis lychnitis is a mostly endemic species of the Iberian Peninsula that frequently hybridizes with the narrow endemic P. crinita subsp. malacitana in southern Spain. Whenever they coexist they form homoploid hybrid zones. Unlike hybridization at the polyploid level, the process of hybridization at the homoploid level is much less well known. METHODS AND RESULTS: In this study we report the development of 22 microsatellite markers through next-generation sequencing technologies for P. lychnitis. We characterize the genetic diversity for two populations of this species for the 10 markers that resulted to be polymorphic. Further, we check the transferability of these polymorphic markers to one population of P. crinita subsp. malacitana to verify the potential use of these markers for hybridization studies. The values of expected heterozygosity for P. lychnitis were higher than in P. crinita subsp. malacitana, and the three analyzed populations displayed negative values for the inbreeding coefficient which is compatible with the frequent instances of hybridization and introgression between species. CONCLUSIONS: This set of polymorphic markers are useful for further studies aiming at a deeper understanding of the homoploid hybrid process between these species. Additionally, this is the first panel of microsatellite markers developed for the genus Phlomis, a genus very rich in endemic species and with medicinal properties that could benefit from the use of these new markers.

Chemical profiling, antiviral and antiproliferative activities of the essential oil of Phlomis aurea Decne grown in Egypt.

Here, we investigated the chemical composition of the edible Phlomis aurea oil and its anticancer potential on three human cancer cell lines, as well as its antiviral activity against Herpes simplex-1 (HSV-1). Exploring Phlomis aurea Decne essential oil by gas chromatography coupled with mass spectrometry (GC/MS) revealed the presence of four major components: germacrene D (51.56%), trans-ß-farnesene (11.36%), α-pinene (22.96%) & limonene (6.26%). An antiproliferative effect, as determined by the MTT assay, against human hepatic, breast and colon cancer cell lines, manifested IC50 values of 10.14, 328.02, & 628.43 µg mL-1, respectively. Cytotoxicity assay of the Phlomis oil against Vero cell lines revealed a safe profile within the range of 50 µg ml-1. Phlomis essential oil induced the apoptosis of HepG2 cells through increasing cell accumulation in sub G1 & G2/M phases, decreasing both S & G0/G1 phases of the cell cycle, triggering both caspases-3 &-9, and inhibiting cyclin dependent kinase-2 (CDK2). The antiviral activity of the oil against HSV-1 was investigated using the plaque reduction assay, which showed 80% of virus inhibition. Moreover, the molecular docking in silico study of the four major chemical constituents of the oil at the CDK2 binding site demonstrated marked interactions with the ATP-binding site residues through alkyl & Pi-alkyl interactions. Cell cycle distribution of HepG2 cells was studied using flow cytometry to highlight the apoptotic mechanistic approaches by measuring caspases-3 &-9 and CDK2 activities. Thus, the edible Phlomis oil can be regarded as a candidate for in vivo studies to prove that it is a promising natural antiviral/anticancer agent.

Anti-proliferative effects of Ziziphus spina-christi and Phlomis russeliana leaf extracts on HEK293 and MCF-7 Cell Lines and Evaluation of Bax and Bcl-2 Genes Expression Level in MCF-7 Cells.

To investigate the effects of Phlomis russeliana and Ziziphus spina-christi leaf extracts on apoptosis in breast cancer MCF-7 cells. Cell lines were divided into a control group and the groups exposed to 0.001, 0.01, 0.1, 1, and 10 mg/ml of Ziziphus spina-christi and Phlomis russeliana leaf extracts. Cell viability was quantified by the MTT assay. The expression of Bax and Bcl-2 genes was evaluated by Real-time PCR analysis. Statistical analysis was performed using ANOVA. HEK293 cell viability significantly increased in the groups exposed to 0.001, 0.01, and 0.1 mg/ml of Z.christi leaf extract and decreased in the group exposed to 10 mg/ml of P.russeliana leaf extract. MCF-7 cells viability significantly decreased in the groups exposed to 0.001, 0.01, 0.1, 1 and 10 mg/ml of Z.christi leaf extract and increased in the groups exposed to 0.001 and 0.01 mg/ml of P.russeliana leaf extract. The exposure of MCF-7 cells to 1 and 10 mg/ml of P.russeliana leaf extract also led to a significant decrease in cell viability. The cytotoxic effect of Z.christi was higher than P.russeliana leaf extracts on MCF7 cells.  1 mg/ml of Z.christi leaf extracts also significantly increased the expression level of Bax and Bcl-2 genes in MCF7 cells. Bcl-2 gene expression significantly increased in the group exposed to 10 mg/ml of P.ruseliana leaf extract.Despite P.russeliana leaf extract, lower Z.christi leaf extract concentrations inhibited MCF-7 cells proliferation. Ziziphus spina-christi and phlomis russeliana leaf extracts mechanism of action has occurred through the Bax-independent apoptotic pathway on MCF-7 cells.

Chemical profiling, antimicrobial, anti-enzymatic, and cytotoxic properties of Phlomis fruticosa L.

Structural diversity of biologically active compounds identified in plants after many years of storage is rarely reported in literature. Herein, we studied chemical profile and biological activities of Phlomis fruticosa L. after plant material storage for 20 years. Chemical analyzes were performed by UHPLC-LTQ-Orbitrap/MS, and revealed presence of 44 compounds: including 13 phenolic acids, 9 phenylethanoids, 20 flavonoids and 2 phenolic related compounds (a phenolic acid derivative and an aliphatic alcohol). The extract showed antimicrobial activity, being the most potent against Aspergillus fumigatus with minimum inhibitory concentration of 0.31 mg/mL. Also, the extract was able to inhibit biofilm formed by Candida species and to inhibit biofilm formation by Staphylococcus aureus. Obtained results revealed that the extract has potential to interfere with the cell membrane permeability of Candida albicans and to suppress production of virulence factor staphyloxanthin in S. aureus. Furthermore, the extract inhibited the activity of α-amylase which is one of the therapeutic targets for diabetes type II. Also, the antiproliferative effect of the extract was demonstrated on human cancer cell lines, while the extract did not exhibit any cytotoxic effect on primary human cells. Based on the obtained results, P. fruticosa could be an interesting source of biologically active compounds even after long term storage.

Chemical composition and antioxidant activity of Phlomis leucophracta, an endemic species from Turkey.

The chemical composition, and antioxidant activity of the essential oils of the endemic Phlomis leucophracta P. H. Davis et Hub.-Mor. was investigated. The major compounds of the essential oil were linalool (36.4%), spathulenol (8.4%) and caryophyllene oxide (8.4%). The composition of the oil differs with published data, suggesting other chemotype. Moreover, the oil of this species was analysed for its antioxidant activity for the first time and results indicate it possess strong antioxidant activity comparable with already known antioxidants, such as ascorbic acid, BHT, or Trolox. Presented results suggest that this endemic species has strong potential to be used in food and pharmacological industries, and therefore they need to be further investigated.

Evaluating antimicrobial and antioxidant capacity of endemic Phlomis russeliana from Turkey and its antiproliferative effect on Human Caco-2 Cell Lines.

In this study, the antimicrobial, antioxidant and antitumor activity of ethanol extracts obtained from Phlomis russeliana (Sims.) Lag. ex Benth. (Lamiaceae) were evaluated. Disc diffusion and microdilution methods were used to test the extracts for antimicrobial activity against seven bacteria strains (Bacillus cereus ATCC 7064, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 6538P, Escherichia coli ATCC 10538, Proteus vulgaris ATCC 6899, Salmonella typhimurium CCM 5445 and Pseudomonas aeruginosa ATCC 27853) and four yeast strains (Kluyveromyces fragilis ATCC 8608, Rhodotorula rubra ATCC 70403, Debaryomyces hansenii DSM 70238 and Candida albicans ATCC 10239). Notably, they were more effective against the yeast strains than the bacterial strains. Of the yeast cultures, D. hanseii was among the most susceptible, having an inhibition zone of 16.2 mm with minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) of 64(128)µg/ml, respectively. For cytotoxic determination, Caco-2 cells were cultured as per ATCC protocol, and were treated with log concentrations (5-80 mg/ml) of P. russeliana. The potency of cell growth inhibition for each extract was expressed as an IC50 value. Moreover, oxidant capacity was evaluated via TOC assay. This product induced antiproliferative activity of 31.33% at 40 mg/ml and 20.96% at 80 mg/ml, without toxic effects on cells, although the oxidant capacity was decreased to 27.06 ± 0.7 nm in the 80 mg/ml-applied group compared to 47.9 ± 1.8 nm in the untreated one. Advanced pharmacological studies are needed to further evaluate P. russeliana for distinctive features.

Chromatographic analysis, antimicrobial and insecticidal activities of the essential oil of Phlomis floccosa D. Don.

The present study was conducted to investigate for the first time the chemical composition and insecticidal and antimicrobial properties of Phlomis floccosa D. Don. essential oil isolated by hydro-distillation of its aerial parts. Qualitative and quantitative analyses of the essential oil on the basis of gas chromatography and mass spectrometry (GC/FID and GC/MS) revealed the presence of 59 organic volatiles representing 86.9% of the total constituents. The major compounds were germacrene-D (19.7%) followed by ß-caryophyllene (15.5%), caryophyllene oxide (8.3%), hexadecenoic acid (7.9%) and carvacrol (6.1%). The broth microdilution method was conducted to test the antimicrobial activity of the essential oil against five pathogens: Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica, Staphylococcus aureus, Bacillus subtilis and the yeast Candida albicans. The oil of P. floccosa D. Don. shows promising activity against the Gram-positive bacteria S. aureus and B. subtilis and the yeast C. albicans (minimum inhibitory concentration = 625 µg.mL-1 ). The effectiveness of the essential oil was assessed on both Tribolium confusum and Sitophilus zeamais pest insects and it was more pronounced against T. confusum, showing a moderate mortality of 47.5%.

Recent Updates on the Phytochemistry and Pharmacological Properties of Phlomis viscosa Poiret.

Crude ethanolic extracts from Phlomis viscosa Poiret leaves from the Judea region (Israel) are renowned for their remarkable geroprotective properties: anti-inflammatory, anti-diabetic, and anti-cancer. A phytochemical investigation carried out in this study revealed that the tested plant might belong to a particular distinct chemotype because its phytochemicals are different from compounds that were mentioned in the literature. Among the compounds identified by us was diosmin, the synthetic derivatives of which were further obtained and investigated. In particular, activities of the isolated compounds and synthesized diosmin derivatives were assessed. Our results revealed that the following compounds significantly lessened secretion of some pro-inflammatory cytokines: diosmin, himachala-2-diene, and 5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl) chromen-4-one. In addition, diosmin, synthesized diosmin derivatives, and some identified terpenes were found to have anti-diabetic activities. A significant anti-cancer effect of the whole extract on U-87 (human glioblastoma carcinoma cells line) and MCF7 (human breast carcinoma cell line) was also demonstrated, and it was better than that of DOX (doxorubicin). Collectively, the results obtained in the in vitro models suggest a wide spectrum of beneficial bioactivities of the extract and its active compounds.

Investigation of the synergistic effects of paclitaxel and herbal substances and endemic plant extracts on cell cycle and apoptosis signal pathways in prostate cancer cell lines.

Paclitaxel, which isolated from Taxus brevifolia, is recently started to be used against prostate cancer treatment and it is a very effective compound against cancer. In this study, we aimed to test the synergistic effect of two plant active compounds (sulphoraphane (SFN) and silymarin (SILY)) and several endemic plant species from Turkey (such as Phlomis leucophracta, Rubia davisiana, Alkanna tinctoria), which are known to have anticarcinogenic effect on androgen-independent PC3 and DU145, and androgen-dependent VCaP prostate cancer cell lines, with paclitaxel on the expression of cell cycle signaling and apoptosis regulator genes. Herbal substances and endemic herbal extracts were combined with Paclitaxel drug. IC50 doses were identified as real-time online. The most effective synergistic doses were determined according to isobologram analysis. The apoptotic effects of effective combined doses were evaluated by TUNEL, Annexin V, and JC-1 methods. Apoptotic and/or cell cycle arrest effects of confirmed combined doses on the expression of genes in these pathways were assessed by real-time online. Endemic plant extracts (Alkanna tinctoria, Phlomis leucophracta and Rubia davisiana, IC50 < 220 µg/ml) and herbal substances (SILY, and SFN IC50 < 130 µM) indicated antiproliferative and apoptotic effects in prostate cancer cell lines. They testified to the synergistic effect of paclitaxel with endemic plant extracts (Combination Index CI, ED50 < 0.41). The combinations, which indicate the synergistic effect was increased to the Bax/Bcl­2 ratio by suppressing Bcl­2 gene expression into the prostate cancer cell lines. Besides, they increased the expression of TNFRSF10A, TNFRSF1A, CHEK1, CDKN1A, CDKN2B, CDK8, CDKN3 and CASP14 and decreased BAD, CDK5RAP1, CDC20, cyclin H, CDK5RAP1, CDC20. The effective doses of paclitaxel were reduced and G2/M arrest was induced by the endemic plant extracts and herbal substances that indicate a synergistic effect with paclitaxel. By using different combination of herbal extracts or active substances with paclitaxel, more economical and efficient treatment strategies can be developed.

De novo assembly of Phlomis purpurea after challenging with Phytophthora cinnamomi.

BACKGROUND: Phlomis plants are a source of biological active substances with potential applications in the control of phytopathogens. Phlomis purpurea (Lamiaceae) is autochthonous of southern Iberian Peninsula and Morocco and was found to be resistant to Phytophthora cinnamomi. Phlomis purpurea has revealed antagonistic effect in the rhizosphere of Quercus suber and Q. ilex against P. cinnamomi. Phlomis purpurea roots produce bioactive compounds exhibiting antitumor and anti-Phytophthora activities with potential to protect susceptible plants. Although these important capacities of P. purpurea have been demonstrated, there is no transcriptomic or genomic information available in public databases that could bring insights on the genes underlying this anti-oomycete activity. RESULTS: Using Illumina technology we obtained a de novo assembly of P. purpurea transcriptome and differential transcript abundance to identify putative defence related genes in challenged versus non-challenged plants. A total of 1,272,600,000 reads from 18 cDNA libraries were merged and assembled into 215,739 transcript contigs. BLASTX alignment to Nr NCBI database identified 124,386 unique annotated transcripts (57.7%) with significant hits. Functional annotation identified 83,550 out of 124,386 unique transcripts, which were mapped to 141 pathways. 39% of unigenes were assigned GO terms. Their functions cover biological processes, cellular component and molecular functions. Genes associated with response to stimuli, cellular and primary metabolic processes, catalytic and transporter functions were among those identified. Differential transcript abundance analysis using DESeq revealed significant differences among libraries depending on post-challenge times. Comparative cyto-histological studies of P. purpurea roots challenged with P. cinnamomi zoospores and controls revealed specific morphological features (exodermal strips and epi-cuticular layer), that may provide a constitutive efficient barrier against pathogen penetration. Genes involved in cutin biosynthesis and in exodermal Casparian strips formation were up-regulated. CONCLUSIONS: The de novo assembly of transcriptome using short reads for a non-model plant, P. purpurea, revealed many unique transcripts useful for further gene expression, biological function, genomics and functional genomics studies. The data presented suggest a combination of a constitutive resistance and an increased transcriptional response from P. purpurea when challenged with the pathogen. This knowledge opens new perspectives for the understanding of defence responses underlying pathogenic oomycete/plant interaction upon challenge with P. cinnamomi.

Variation in Chemical Composition and Antibacterial Activity of the Essential Oil of Wild Populations of Phlomis olivieri.

Various species of the genus Phlomis have been reported to produce metabolites demonstrating significant pharmacological efficiency. In this study, the essential oils from twelve populations of Phlomis olivieri collected from natural habitats were investigated for their chemical components. The hydrodistillated essential oil analyzed by GC-FID and GC/MS. Analyses revealed 27 compounds, constituting 90.52 - 98.51% of the essential oils. Results indicated that the major components of the essential oils from various populations of P. olivieri were germacrene D (26.54 - 56.41%), bicyclogermacrene (6.38 - 30.55%), ß-caryophyllene (5.32 - 24.52%) and α-pinene (1.29 - 15.53%). Principal component analyses (PCA) was used to identify any geographical variations in essential oil composition. Notably, three groups of Iranian P. olivieri populations were determined according to the major compounds. Results of the in vitro antibacterial activity indicated that P. olivieri essential oils showed good inhibitory activities against bacteria, especially Bacillus subtilis. The results of this study gave new insights for cultivation and industrial uses of P. olivieri in Iran.

Structure, anti-Phytophthora and anti-tumor activities of a nortriterpenoid from the rhizome of Phlomis purpurea (Lamiaceae).

To investigate bioactive compounds potentially involved in the biotic interactions exhibited by Phlomis purpurea (Lamiaceae) in rhizospheres infested with Phytophthora cinnamomi, the plant rhizome was chemically analysed. The nortriterpenoid (17S)-2α,3α,11α,23,24-pentahydroxy-19(18 â†’ 17)-abeo-28-norolean-12-en-18-one, was isolated and its structure was elucidated by comprehensive spectroscopic analysis, chiefly using 2D NMR experiments, and X-ray analysis. It was shown to be exuded by roots and to exhibit anti-Phytophthora and antitumor activities.

Biomimetic synthesis of antimicrobial silver nanoparticles using in vitro-propagated plantlets of a medicinally important endangered species: Phlomis bracteosa.

In vitro-derived cultures of plants offer a great potential for rapid biosynthesis of chemical-free antimicrobial silver nanoparticles (AgNPs) by enhancing their phytochemical reducing potential. Here, we developed an efficient protocol for in vitro micropropagation of a high-value endangered medicinal plant species, Phlomis bracteosa, in order to explore its biogenic potential in biomimetic synthesis of antimicrobial AgNPs. Murashige and Skoog medium supplemented with 2.0 mg/L thidiazuron was found to be more efficient in inducing optimum in vitro shoot regeneration (78%±4.09%), and 2.0 mg/L indole-3-butyric acid was used for maximum root induction (86%±4.457%). Antimicrobial AgNPs were successfully synthesized by using aqueous extract (rich in total phenolics and flavonoids content) of in vitro derived plantlets of P. bracteosa. Ultraviolet-visible spectroscopy of synthesized AgNPs showed characteristic surface plasmon band in the range of 420-429 nm. The crystallinity, size, and shape of the AgNPs were characterized by X-ray diffraction and scanning electron microscopy. Face-centered cubic AgNPs of almost uniform spherical size (22.41 nm) were synthesized within a short time (1 hour) at room temperature. Fourier-transform infrared spectroscopy revealed that the polyphenols were mainly responsible for reduction and capping of synthesized AgNPs. Energy dispersive X-ray analysis further endorsed the presence of elemental silver in synthesized AgNPs. These biosynthesized AgNPs displayed significantly higher bactericidal activity against multiple drug-resistant human pathogens. The present work highlighted the potent role of in vitro-derived plantlets of P. bracteosa for feasible biosynthesis of antimicrobial AgNPs, which can be used as nanomedicines in many biomedical applications.

[Molecular Identification on Three Medicinal Phlomis Species Produced in Yunnan Province Based on ITS and matK].

Objective: To analyze ITS region and matK gene sequences of three medicinal Phlomis plants,in order to provide molecular basis for identifying and protecting their wild resources. Methods: PCR and sequencing were conducted on Phlomis likiangensis,Phlomis melanantha and Phlomis betonicoides wild populations by primers pairs ITS4 / ITS5 and matKXF / matK5 R. Results: The smallest inter-K2 P genetic distance was further than the largest intra-K2 P genetic distance in Phlomis likiangensis, Phlomis melanantha and Phlomis betonicoides. Different samples of three medicinal Phlomis plants were gathered together and could be distinguished from other exogenous species by Neighbor-Joining( NJ) tree. Phlomis likiangensis, Phlomis melanantha and Phlomis betonicoides had three, three and one sites on ITS2 for their effective identification, and had three,three and three sites on ITS1 for their effective identification respectively. Phlomis betonicoides had three sites on matK for its effective identification, while Phlomis likiangensis or Phlomis melanantha needed multiple sites for their effective identification. Conclusion: The study implies that ITS1,ITS2 and matK fragments could be used for molecular identification of Phlomis likiangensis, Phlomis melanantha and Phlomis betonicoides.

T cell regulation by Phlomis lanata protein extracts in mice.

CONTEXT: Phytopharmacology is a complex but very promising research area. The different plant parts and extraction methods may result in opposed effects. Phlomis species have been reported for both anti-inflammatory and tonic properties. OBJECTIVE: The effect of Phlomis lanata Willd. (Lamiaceae) protein extracts on immune cell reactivity was studied in the experimental mouse model. MATERIALS AND METHODS: Protein extracts from P. lanata aerial parts were fractionated by Q-sepharose ion-exchange chromatography and applied to whole spleen cells or T-cell subsets at 5 µg/ml. Cell growth and cytokine production were evaluated after 4 and 2 d of culture using (3)H-thymidine-uptake and ELISA techniques, respectively. RESULTS: Among the protein fractions tested, column wash proteins (W1) and the fraction eluted using 600 mM NaCl (F6) reduced by 76% and increased by 78% spleen cell proliferation, respectively. W1 suppressed proliferation of effector T-cells, but stimulated the growth of suppressor/regulatory cells by 62-148%. Although W1 stimulated IL-2 and IL-10 production from total spleen cells, it significantly increased IL-10 (50%) and reduced IL-2 (30-50%) production from T-cells, while TNF-α release was enhanced in CD25(+)CD4(+) by 92% and reduced by 50% in CD25(+)CD8(+) cells. F6 stimulated whole spleen cell growth, reduced proliferation of CD8(+) and CD25(+) cells by approximately 50%, while decreasing by 60-80% TNF-α production from CD25(-) and CD25(+)CD8(+) cells. DISCUSSION AND CONCLUSION: The suppressive activity of W1 could be attributed to IL-10 and TNF-α, while the stimulatory effect of F6 could be attributed to the inhibition of T-regulatory cells. In the same plant, coexisting protein fractions induce both immunostimulatory and immunosuppressive activities.