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Plant endophytes deployment at the field level affects the productivity of agricultural products. Savory Satureja khuzestanica Jamzad is a valuable medicinal plant native to Iran. In this study, in order to improve the quantitative and qualitative yield of the plant, the development of a suitable method for inoculation of native fungal endophytes to S. khuzestanica is surveyed. Three fungal endophytes (P.I.S.7, D.2.F.1 and D.1.S.1), isolated from savory plants, were inoculated on S. khuzestanica using four different methods (stem injection, foliar spray, soil drench and adding fungi to the soil in granular form) and their impact on plant's growth and its physiological response was analyzed. All inoculation methods, except adding granular form of fungi to the soil, resulted in endophytic colonization of the savory tissues. However, P.I.S.7 isolate was not established in these plants using any inoculated methods. Extent of fungal colonization showed a significant difference with the inoculation method, such that foliar spray had the highest colonization level, followed by stem injection and root drench. The highest rate of fungal colonization belonged to D.1.S.1. isolate. Spraying leaves and stem injection with endophytic fungi, especially in the case of D.1.S.1, significantly enhanced the morphological and physiological characteristics of host plants and the percentage of essential oil yields in these plants. All in all, using the D.1.S.1 isolate and foliar spray method led to savory plants' improved quantitative and qualitative product.

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In this study, the aerial parts of Pimpinella puberula were collected from Ramhormoz and Mashhad (Khuzestan and Khorasan Provinces) at the vegetative, flowering and seeding stages. Essential oils from the whole aerial parts as well as stem/leaf, inflorescence, unripe and ripe seeds, were isolated by hydro-distillation. The yields of essential oil obtained from the Ramhormoz samples were 0.49%, 0.31%, 3.81%, 6.01% and 1.80% w/w, and from those from Mashhad were 0.96%, 0.87%, 3.59%, 6.94% and 4.96% w/w, respectively. The oils from different parts of plant were also analyzed by GC and GC/MS. Limonene was the major constituent in all the oils (21.7%-82.4%), followed by pregeijerene (14.6%-55.4%) and geijerene (7.2%- 11.7%). Methyl eugenol and elemicine, however were only found in the Ramhormoz oil samples. The antimicrobial activity of the oils was determined using the disk diffusion method against Gram positive bacteria (Bacillus subtilis, Bacillus cereus, Micrococcus luteus and Staphylococcus aureus), Gram negative bacteria (Yersinia entrocolitica, Klebsiella oxytoca, Serratia marcescens, Escherichia coli and Pseudomonas aeruginosa) and yeast (Candida albicans). Results showed a significant difference between Gram positive and Gram negative bacteria in their susceptibility to the oil, although Gram positive bacteria were more susceptible to the antimicrobial activity of P. puberula oil. In addition, the antimicrobial activity of samples collected from Ramhormoz were more than of those from Mashhad.

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Objective: Despite toxic effects of some essential oils, their use is not under control. With a view to increasing trend of utilisation of herbal products, some biological aspects of Thymus daenensis are repoted here for the first time. Materials and Methods: Antimicrobial properties using disk diffusion and dilution tests, nitric oxide radical scavenging by Marcocci et al method and cytotoxic properties employing dimethylthiazolyl diphenyltetrazolium bromide reduction test were carried out with Thymus daenensis and commercial Thyme essential oils and their main chemical compound, thymol. Results: The microbial sensitivity to the oils were in Candida albicans> E.coli> S.aureus> P. aeruginosa order. The minimum inhibitory and microbicidal concentrations were in the range of 0.04-10mg/ml. Nitric oxide radical scavenging was dose dependent with an IC50 of 5, 75, 863 µg, and total phenolics of 644.07±6.79, 16.94±2.55, 10.33±2.31µg Gallic acid equivalent per mg sample and total flavonoid content of 73.51±1.34, 0.56±0.02, 0.21±0.09 mg Catechin equivalent per gram T.daenensis oil, commercial thyme oil and thymol respectively. The concentrations from T.daenensis oil, commercial thyme oil and thymol required to exert 50% fatal effect (IC50) on healthy human normal lymphocytes and Hela cells were 1455, 12.10, 2867 and 4.95, 3.61, 1730 µg respectively. Conclusion: T. daenensis with its good antimicrobial property can prevent formation of toxic reactive oxygen species and as a good antioxidant, it can directly scavenge NO and O2−. With a view to cancerous cells killing properties of the oils at their lowest concentrations without fatal effect on normal healthy cells, feasibility of their application in combating cancerous cells may be promising.

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Many parameters affect essential oil yield and chemical composition of aromatic plants. For obtaining the highest quantity and quality of essential oil, it is necessary to know the proper methods of drying and distillation. The aim of this research was to investigate the influence of drying and extraction methods on the yield and chemical composition of the essential oil of Eucalyptus sargentii. The fresh leaves of Eucalyptus sargentii, cultivated in Kashan (central region of Iran), were collected in the middle of spring and dried by five different drying methods: sun-drying, shade-drying, and oven-drying at 30, 40 and 50°C. The essential oils of every treatment were obtained by hydro-distillation in three replication. In addition, the essential oil of shade-dried sample was obtained by two other distillation methods, namely, water- and steam-distillation and direct steam-distillation. The oils were analyzed by capillary GC and GC-MS. Statistical analysis showed significant difference between oil yield (w/w) of the shade-dried sample (3.39%) compared to oven-dried at 40°C (2.92%), sun-dried (2.66%), oven-dried at 30°C (2.59%) and oven-dried at 50°C (2.30%). The oil content of the shade-dried sample obtained by hydro-distillation (3.39%) was higher than those of the water and steam distillation (2.89%) and steam distillation (1.35%). Twenty-three components were identified in the oil of E. sargentii in the different drying methods, including 1,8-cineole (57.9-65.8%) and α-pinene (11.3-28.3%) as main components. Twenty-four compounds were characterized in the oils of different distillation methods, including 1,8-cineole (61.2-66.6%) and α-pinene (19.7-28.3%) as the major compounds. Among the different drying methods, shade-dried samples produced the highest oil yield and 1,8-cineole content, while in different distillation methods, hydro-distillation produced the highest oil yield, but the highest percentage of 1,8-cineole was obtained by water and steam distillation.

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Satureja sahendica Bornm is an endemic, native, and aromatic plant in Iran, with thymol being its main and major volatile component, which is used in food and pharmaceutical industries. Increasing the oil yield and phenolic compounds in essential oil leads to increased medicinal effects. In this study, the effects of mycorrhizal fungi (Glomus mosseae, G. intraradices, and combination of G. mosseae and G. intraradices) and different levels of water stress [D₁= Control (no water stress), D₂= No irrigation during stem elongation till blooming, D₃= No irrigation at the blooming up to start of flowering, and D₄= No irrigation at 50% flowering up to full flowering] were studied on S. sahendica oil, in two years. When the seedlings were transferred to the field, 10 g of mycorrhiza fertilizer (containing mycorrhizal fungus spores) were added to the rhizosphere of each seedling. The plants were harvested at full flowering and the essential oils were obtained by hydro-distillation and analyzed by GC and GC/MS. The highest amount of oil yield (66.13 kg ha^-1) was obtained using G. intraradices in non-stress conditions in the second year. The highest percentage of thymol was obtained in G. mosseae×control in the first year (74.59%), whereas in the second year, the highest amount of thymol was in G. mosseae×D₄ (61.97%). This study showed that with the use of mycorrhizal fungi, even in conditions of water shortage, S. sahendica produced more essential oil with a higher percentage of thymol. With the use of mycorrhizal fungi, the essential oil can easily be increased and the number of compounds changes.
 

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The genus Nepeta is one of the largest genera in the Lamiaceae family, and Iran is one of the main centers of origin of this genus. The genetic diversity estimation of the genus Nepeta germplasm provides a basis for future biodiversity conservation efforts and also for the selection of high-productive genotype(s) for the field production improvement. In this work, eleven RAPD primers with suitable banding pattern and prominent polymorphism were used for the estimation of infraspecific genetic diversity of N. kotschyi based on 21 populations. The study included 19 and 2 populations belonging to var. persica and var. kotschyi, respectively. Eleven primers amplified totally 225 scorable RAPD loci, 204 of which were polymorphic. The average number of bands per primer was 20.5, 18.5 of which were polymorphic. Primers OPF 05, OPB 15, OPT 14, OPO 07, and OPF 14 were the most powerful for the detection of the genetic diversity across the samples. Cluster analysis showed six genotypic groups. N. kotschyi var. kotschyi populations were placed in a group separated from the samples belonging to N. kotschyi var. persica. The same analysis showed that the genetic diversity pattern corresponds to the geographical distribution of the populations. Of the total variation in the studied germplasm, 32% was related to inter-populations and 68% to intra-populations variation. The results indicated the high potential of RAPD markers to resolve the status of the studied genotypes in regard to inter- and intra-populations diversity and to diversify N. kotschyi varieties previously resolved by morphometric methods.

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Qualitative and quantitative variations in the essential oil of wild growing Origanum vulgare L. ssp. gracile plants were studied in response to different phenological stages (pre, full and post-flowering). The essential oil of air-dried leaves was isolated by water distillation using a Clevenger-type apparatus and was analyzed by Gas Chromatography (GC) and Gas Chromatography–Mass Spectrometry (GC/MS). The highest (1.87%) and the lowest (1.01%) essential oil content were obtained from post-flowering and pre-flowering stages, respectively. In total, 24 components were identified and quantified in three phenological stages representing 96.75, 97.63, and 98.59% of the oil, respectively. Carvacrol (46.62, 46.5 and 27.6%), ρ-cymene (7.76, 13.54 and 37.08%) and γ-terpinene (21.54, 13.91 and 6.82%) were the main constituents of essential oils in pre, full, and post-flowering stages, respectively. Oxygenated monoterpenes (43.35-61.32%) and monoterpene hydrocarbons (30.81-48.02%) were the main classes of identified compounds in three essential oils. According to the findings of this research, the post-flowering stage can be considered as the most appropriate time for obtaining the highest essential oil content, but to achieve the highest rate of phenolic compounds, the pre-flowering and full-flowering stages can be recommended.

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Achillea tenuifolia Lam. belongs to the Asteraceae family that is one of the most popular aromatic plants in Iran with great variation in both morphology and essential oil components. The aim of this study was determination of essential oil content and composition of A. tenuifolia populations in field condition. Seeds of 17 populations of A. tenuifolia collected from their natural habitats were cultivated under the same environmental conditions in the research farm of Alborz Research Station, Karaj, Iran. The aerial parts of the plants were collected at full flowering stage and dried in shade (room temperature) and their essential oils were obtained by hydro-distillation. The oils were analyzed by GC and GC/MS. Based on the results, the oil yield varied from 0.16 to 1.59% (w/w dried weight). Populations from Divandareh with 1.59% and Khoy2 with 0.16% had the highest and lowest oil yield, respectively. According to the cluster analysis, the populations were placed in two clusters. Germacrene D was the main compound in all oils, but the essential oils of populations in cluster 1 contained higher amount of germacrene D (up to 64.5% in Semnan population). α-Humulene (up to 15%) and 1,8-cineole (up to 11.7%) were the other major components in the oil of cluster 1 populations. The lower amounts of these three compounds (in addition to E-β-farnesene and piperitone) and higher amount of more volatile compounds like p-cymene, β-phellandrene, camphor, and α-thujone and presence or absence of other minor compounds placed the other populations in cluster 2. Each cluster divided in two groups because of different percentages of some component such as cubenol, viridiflorol, methyl hexadecanoate and phytol. Therefore, based on the demand for processing, the proper population can be chosen for vast cultivation.