Journal of Medicinal Plants and By-products
Volume:5 Issue: 2, Winter and Spring 2016

Callus induction responses and regeneration through callus-mediated SE were studied from evening primrose (Oenothera biennis L.) as an important medicinal plant of Onagraceae mainly known for its gamma-linoleic acid (GLA) content. The effects of cytokinins, 6-Benzylaminopurine (BAP) and N-(2-furfurylamino)1-H-purine-6amine [Kinetin (Kin)] and their concentrations (0, 0.5, 1.0 and 1.5 mg/l) in combinations with 2,4dichlorophenoxyacetic acid (2, 4-D) concentrations (0.25, 0.75 mg/l) on callus induction and SE for three explant types (petiole, leaf and epical bud) of "Shiraz" variety were separately evaluated. In all types of explants, the highest callus fresh weights belonged to 1 mg/l BAP or Kin in combinations with 2, 4-D (either 0.25 mg/l or 0.75 mg/l). The maximum fresh weight of callus was obtained from leaf explants plated on culture medium containing of 0.25 mg/l 2, 4-D and 1 mg/l Kin. The maximum The maximum number of embryos was achieved from leaf explants related to 0.75 mg/l 2, 4-D and 1 mg/l Kin. plantlets were successfully raised from in vitro developed embryos. Efficient plant regeneration via SE may provide a reliable system for studying the molecular mechanism of SE and a route for the genetic transformation of evening primrose.

In this research, effect of vermicompost and plant growth promoting rhizobacteria (PGPR) on physiological traits of lemon balm under drought stress condition was investigated in year 2012 at Karaj Research Institute of Forest and Rangelands. Experiment was conducted as split-split plot in a randomized complete block design with three replications. irrigation (normal: irrigated to field capacity; drought stress: watered to 60% of field capacity) was considered as main plot, vermicompost fertilizer (consuming 0, 5 and 10 t ha-1) as split plot and biofertilizers (Pseudomonas fluorescent, Azotobacter chrococum̺Ⲧ⫢陙 brasilense, Azotobacter Azospirillum Pseudomonas and no fertilizer) as split-split plot. Results showed that water stress significantly increased essential oil content, while decreased essential oil yield, total chlorophyll, cell membrane stability, relative water content and proline. Only relative water content and total chlorophyll were positively affected by biofertilizer application. Essential oil content, essential oil yield, total chlorophyll, cell membrane stability, relative water content and proline were significantly increased by vermicompost application, especially, relative water content increased with an increase in amount of vermicompost application, but there were strong interactions with irrigation. These interactions are important in practical use of vermicompost and biofertilizers.

Two field and laboratory experiments were conducted to investigate the effects of superabsorbent polymer (SAP) and irrigation management on seed and essential oil yields of Cumin, as well as the impact of water quality on water holding capacity of SAP. Salinity had a negative effect on the amount of water absorbed by SAP (335 and 59 g H2O per g SAP, for distilled water and solution of 0.5% NaCl, respectively). SAP application (30 kg.ha-1) along with three times irrigation at sowing, flowering and seed filling stages increased the amounts of seed and essential oil yields by 2.79 and 3.05 times, compared to control. Positive effects of SAP were related to enhancement of soil water holding capacity (120 gr irrigation water per gr SAP), leaf area duration (one week) and subsequently grain filling period.

The Achillea L. genus has a wide distributional range, and the differences in oil composition may be affected by different environmental factors such as soil mineral fertilization, the climate conditions and the culture site. To evaluate the effect of temperate climate on content and chemical combination of essential oil of Achillea filipendulina Lam, A. arabica Kotschy and A. eriophora DC., this study was carried out. Essential oils were analyzed by GC and GC/MS. Yield of essential oil of A. filipendulina, A. arabica and A. eriophora cultivated was 0.9, 2.25 and 2.3%, respectively .The major components of A. filipendulina were santolina alcohol (37.2%), borneol (12.7%),1,8- cineole (8.7%), germacrene D (6.22%), and camphor (4.09%). In A. arabica, the major compounds were p-cymene (14.61%), piperitone (13.09%), camphor (12.81%) and 1, 8-cineole (11.97%). In A. eriophora, principal compounds were camphor (19.57%), 1, 8-cineole (19.06%), camphene (9.61%) and α-pinene (7.35%). Our results demonstrate variations in the qualitative and quantitative composition in the oils obtained from the aerial parts of the studied species. Also, chemical profile of A. filipendulina was slightly affected under temperate climatic conditions, suggesting its ability and flexibility to be cultivated in temperate zones.

The focus of the present study was to investigate the chemical composition of essential oil of Salvia hydrangea DC. ex Benth. growing wild in Kiasar-Hezarjrib, Iran. In addition, effects of altitude and some soil properties on the essential oil quality and quantity of S. hydrangea was identified. The samples collected from two altitudes (1100 and 2000 m) in blossoming period, were analyzed by hydrodistillation method using a clevenger-type apparatus by GC and GC/MS. Ninety-one compounds were identified, representing 99.79% and 99.95% of the oils from the altitudes of 1100 and 2000 m respectively. The main constituents of the oils in altitude 1100 were 1,8-cineole (12.70%), camphor (12.06%), β-pinen (11.62%), naphthalene (10.83%), α-amorphene (9.30%), bicycleheptan (9.18%) and δ-cadinene (8.25%). In contrast, in altitude of 2000 m the oil of plant species was rich in naphthalene (19.94%), 1,8-cineole (9.45%), camphor (5.71%), α-terpineol (5.11%) and ylangene (4.94%). In both altitudes naphthalene, 1,8-cineole and camphor being the major compounds respectively. Results indicated that the altitude of 2000 m revealed greater quantity. The essential oil compositions showed the highest percentage in the altitude of 1100 m and amount of main and common essential oil compositions was more in the altitude of 1100 m. A high positive correlation was found between the essential oil efficiency and the soil factors of pH, S.P, EC, Mn and Mg in both altitudes while, the common essential oil composition showed negative correlation with these factors. In general, the higher essential oil efficiency and compositions were found in the higher altitude, but percentage of the essential oil compounds was more in the lower altitude therefore, at lower altitudes there are more optimal conditions for taking the S. hydrangea essential oil.

The Lamiaceae family consists of a broad spectrum of medicinal plants involving Marrubium L. genus. Regarding the diverse pharmaceutical uses of the plants belonging to this genus, they can be considered as proper alternatives for chemical drugs having harmful effects. The present work aims to identify and characterize chemical compositions of the essential oils and volatiles from the aerial parts of Marrubium persicum C. A. Mey. as an herbal plant in Iran using classical hydrodistillation. To establish a comprehensive comparison between the traditional techniques and advanced ones, microwave-based extraction techniques namely MAHD as well as SFME have also been utilized. In another part of this project, the profiles related to the volatile fractions from the aerial parts of Marrubium persicum C. A. Mey.have been assessed and compared with the other categories. The main components in the hydrodistillation (HD) method were α-pinene (21.5%), spathulenol (19.5%), α-thujene (17.4%), while the headspace solid-phase microextraction (HS-SPME) profile mainly consisted of β-caryophyllene (14%), eugenol (11.2%) and methyl eugenol (10.2%). On the other hand, using the SFME approach spathulenol (25.4%), α-pinene (17.4%) and germacrene D (9.5%) were found as the most abundant constituents. Moreover, in the MAHD profile caryophyllene oxide (13.1%), δ-elemene (12.4%), camphene (8.5%) were respectively the predominant natural compounds. According to gas chromatographicmass spectrometric determinations, a total of 40 compounds were recognized in the corresponding profiles totally covering 94.6-99.7% of the whole chemical compositions. Sesquiterpene hydrocarbons were recognized as the most frequent groups of natural compounds in the profiles of the advanced approaches, whereas in the traditional one monoterpene hydrocarbons were found to be the dominant constituting group.

The genus Tanacetum L. belongs to the Asteraceae family and Anthemideae tribe is one of the most important medicinal plants that contains 26 species in Iran, 12 of them are endemic. This paper reports the essential oil composition of Tanacetum uniflorum (Fisch. & C.A. Mey. ex DC.) Sch.Bip. growing spontaneously in Salmas (North-West of Iran). Essentail oil extracted by water distillation and steam distillation from Plant on flowers and leaves were collected on July - 2016. The yields of water distillation from leaf were 0.12% and flower were 1.16% (V/W), and by steam distillation from leaf were 0.33% and flower were 0.76% (V/W), respectively, then samples were analyzed by GC and GC/MS. Main components obtained from water distillation from leaf were manoyl oxide 28.87% , α-cadinol 9.7% and γ-eudesmol acetate 4.77% and in flower were methyl pentanoate 88.28%, butyl acetate 5.13% and iso-dihydro carveol acetate 1.19% (V/W), and by steam distillation from leaf were γ- terpinene 17.99%, endo-fenchyl acetate 9.63% and butyl acetate 8.12% and in flower were methyl pentanoate 79.9%, bulnesol 2.70% and (E )-nerolidolol acetate 2.69% obtained, respectively.

Considering great medicinal value of Hyssopus officinalis L. and possibility of its mass production through in vitro culture, two individual experiments was conducted. Effect of various concentrations (0, 2.2, 4.4 and 11 µmol) of TDZ and BAP in combination with 1 µmol of IAA on direct regeneration from nodal explants were assessed. Significant difference between treatments was observed (P≤ 0.01). In BAP treatments, the maximum shoot-buds induction (9 shoot-buds per explant) and shoot regeneration percentage (96.66%) were observed on MS medium fortified with 2.2 and 4.4 µmol BAP in combination with 1 µmol of IAA. In TDZ treatments, the highest regeneration percentage was achieved in MS medium supplemented with TDZ (2.2 µmol) and IAA (1 µmol), and the maximum shoot-buds induction (19.83 shoot-buds per explant) was observed in medium containing 4.4 µmol of TDZ in combination with 1 µmol of IAA. The highest root production frequency (89.5%) was achieved in medium contained 9.84 µmol of IBA. Rooted plants were acclimatized successfully in greenhouse conditions with 100% survival. The protocol described here could be applicable for mass in vitro production of the valuable medicinal plant Hyssopus officinalis L. for its genetic resource conservation as well as pharmaceutical purpose.

In this study the effect of different concentrations of some abiotic elicitors (salicylic acid, jasmonic acid, cadmium chloride and sodium chloride) with different incubation times (3, 6, 9 and 12) on azadiractin production and cell growth in the cell culture of the Iranian native Neem were investigated. Cell growth rate was measured by changes in dry cell weight and it was used as a basis for the treatment of elicitor effect. Azadirachtin was extracted with methanol and measured by high performance liquid chromatography (HPLC). The results showed that the AZ content of cells increased significantly in the cells elicited with different types and concentrations of all used elicitors compared to the control. The cell suspension which treated with 0.75 mM salicylic acid showed the highest Azadirachtin content (0.1268 g/g DCW) 6 days after incubation. The highest dry cell weight, with a significant increase, was observed in the suspension cultures treated with 0.1 mM cadmium chloride 12 days after incubation. However, this study highlighted the new valuable Iranian native Neem genotype and salicylic acid as a powerful elicitor for azadirachtin production. In addition, it was approved the effect of exposure time, individual intrinsic and more presumably combined condition of elicitors and their concentration on plant secondary metabolites production.

Parthenolide is major Sesquiterpene lactones present in Anthemis haussknechtii Boiss. & Reut. (feverfew). This compound has many effects on different disease such as migrain and cancer. Parthenolide was reported from Tanacetum parthenium (L.) Sch.Bip. but other plants of Asteraceae family could contain parthenolide. In this study parthenolide was extracted and identified with two methods, Fourier Transform Infra-Red Spectroscopy (FTIR) and High Performance Liquid Chromatography (HPLC). Breast cancer cell line, MDA-MB-231, was exposed to different concentrations of parthenolide for 24 hours. Half maximal inhibitory concentration (IC50) was evaluated using Methylthiazol Tetrazolium (MTT) test. Based on results, 1000 µg/ml concentration is the minimum lethal dose that kills approximately 50% of cells after 24 hours. The results revealed that the A. haussknechtii parthenolide dramatically decreased survival of cancer cell line by inducing apoptosis. This is the first report of cytotoxicity effect of A. haussknechtii extract on breast cancer cell line.

Otostegia persica (Burm.) Boiss. is the medicinal plant of Lamiaceae family that has long been used traditionally in treatment of severe diarrhea, common cold, flatulence, headache, sore, fever, skin allergies, and blood fat and sugar. This research was conducted in 2013 for one year to identify some ecological characteristics and potential allelopathic of O. persica in Hormozgan province. The climate, geology, geomorphology, land types, associated species, distribution map, growth parameters, soil physical and chemical properties, and the phenology were studied in five regions, including Abmah, Bokhon, Geno, Tangezagh and Bashagard. Also, effects of aqueous extract prepared from leaves of O. persica on 6 species of Brassica oleracea L., Raphanus sativus L., Lepidium sativum L., Coriandrum sativum L., Zinnia elegans L. and Calendula arvensis M. Bieb. were studied. This species is distributed in an altitude of 250-1850 m a.s.l, from the northernmost point of the province to the closest region to the Persian Gulf and from the westernmost point to Bashagerd in the east province. The climate of the study area is arid and hot desert. More than 57.6% of the habitats is mountainous of which 54.6% has limestone formations. The soil texture is sandy loam with pH and EC ranging from 7.42-8.36 and 0.97-1.52, respectively. Aqueous extract prepared from leaves of O. persica inhibited seed germination and decreased speed germination and seedling growth.

Phytochemical constituents, such as total phenol, allicin and pyrovic acid as well as antioxidant activity of thirteen ecotypes of Allium hirtifolium Boiss. from different regions of Iran were evaluated. Average contents of total phenol, allicin and pyrovic acid of bulbs extracts per g fresh weight were 0.647 mg gallic acid equivalent (GAE), 2.127 mg and 84.412 µmol, respectively and antioxidant activity was 63.717%. The ecotypes that exhibited greater contents of studied phytochemicals consequently possessed higher antioxidant activities. Among the analyzed antioxidant enzymes, superoxide dismutase activity was present in the greatest quantity (61.501 U mg-1 protein), followed by ascorbate peroxidase (54.182 U mg-1 protein), polyphenol oxidase (46.219 U mg-1 protein), peroxidase (1.972 U mg-1 protein) and catalase (0.49 U mg-1 protein). Correlation analysis showed the accumulation of A. hirtifolium phytochemicals was associated with mean annual temperatures and precipitation. Cluster analysis on phytomedical characters arranged the ecotypes in five groups. The ecotypes of group D presented by Isfahan ecotypes showed the highest pharmaceutical potential which could be considered in future breeding programs. The ecotype groups were not strictly concordant with their bioclimatic or geographic location, so it can be concluded that the genetic factors as well as environmental factors affected the antioxidant capacity of A. hirtifolium ecotypes.

Studies have shown that oxygenated compounds and TSS/TA are important for beverage and food industries. It seems that Citrus rootstocks have a profound influence on oxygenated compounds and TSS/TA. The aim of this research is to identify rootstock that can synthesize the maximum level of oxygenated compounds and TSS/TA. Peel oil was extracted using mechanical presses and eluted using n-hexane. Finally compounds were analyzed using GC-FID and GC-MS. Total soluble solids were determined using a refractometer. Twenty-seven, twenty-five and twenty-three compounds were identified in Sour orange, Swingle citromelo and Troyer citrang rootstocks, respectively. Limonene (92.87% to 93.16) and myrcene (1.57% to 1.79%) were the main compounds. Among the three rootstocks studied, Swingle citromelo demonstrated the maximum level of oxygenated compounds and TSS/TA. As a result of our research, we can express that the rootstocks can affect the amount ofoxygenated compounds and TSS/TA.

A laboratory trial was conducted to determine whether suspensions of ZnO nanoparticles (ZnO NPs) could interfere with the early growth of fenugreek. This plant species is one of the recommended species by Organization for Economic Cooperation and Development (OECD). Nine concentrations of ZnO NPs (10, 50, 100, 500, 1000, 2000, 3000, 4000, and 5000 mg/L) were prepared in deionized (DI) water (considered as a control). Seed soaking and incubation of seeds in ZnO NPs suspensions were compared. We found that ZnO NPs cannot pass through the seed coat, because neither the seed soaking affect seedling growth nor the germination rate was not affected by ZnO NPs. The root and shoot growth were not affected until 100 and 500 mg/L, respectively, but in concentration more than 100 and 500 mg/L, root and shoot growth negatively were affected. Therefore root growth upon exposure to ZnO NPs was more sensitive than shoot growth.